Ethnomedicinal uses, phytochemistry and pharmacological properties of the genus Boerhavia.

Author’s Accepted Manuscript Ethnomedicinal uses, phytochemistry and pharmacological properties of the genus Boerhavia Kapil S. Patil, Sanjivani R. Bhalsing

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To appear in: Journal of Ethnopharmacology Received date: 12 August 2015 Revised date: 27 January 2016 Accepted date: 31 January 2016 Cite this article as: Kapil S. Patil and Sanjivani R. Bhalsing, Ethnomedicinal uses, phytochemistry and pharmacological properties of the genus Boerhavia Journal of Ethnopharmacology, http://dx.doi.org/10.1016/j.jep.2016.01.042 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting galley proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

Ethnomedicinal uses, phytochemistry and pharmacological properties of the genus Boerhavia Kapil S. Patil and Sanjivani R. Bhalsing* Department of Biotechnology, School of Life Sciences, North Maharashtra University, Jalgaon- 425 001, Maharashtra, India. *E-mail: [email protected] Tele phone: +91 0257 2257421 Fax: 0257-2258403 Abstract: Ethnopharmacological relevance: The genus Boerhavia is widely distributed in tropical, subtropical and temperate regions of the world including Mexico, America, Africa, Asia, Indian Ocean Islands, Pacific Islands and Australia. The genus Boerhavia is extensively used by local peoples and medicinal practitioners for treatments of hepatitis, urinary disorders, gastro intestinal diseases, inflammations, skin problems, infectious diseases and asthma. Present review focused on traditional uses, phytochemistry, pharmacology and toxicology of Boerhavia genus to support potential scope for advance ethnopharmacological study. Materials and methods: Information on the Boerhavia species was collected from classical books on medicinal plants, pharmacopoeias and scientific databases like PubMed, Scopus, GoogleScholar, Web of Science and others. Also scientific literatures based on ethnomedicinal surveys, Ph.D. and M.Sc. dissertations, published papers from Elsevier, Taylor and Francis, Springer, ACS as well as Wiley publishers and reports by government bodies and documentations were assessed. Results: A total of 180 compounds from Boerhavia genus were isolated of which B. diffusa alone shared around 131 compounds and for most of which it is currently an exclusive source. In the genus, phenolic glycosides and flavonoids contribute approximately 97 compounds. These includes eupalitin, rotenoids like boeravinones, coccineons, alkaloid i.e. betanin and punarnavine etc., showing vital pharmaceutical activities such as anticancer, anti-inflammatory, antioxidant and immunomodulatory. Conclusion: Boerhavia is an important genus with wide range of medicinal uses. However, most of the available scientific literatures have lacked relevant doses, duration and positive controls for examining bioefficacy of extracts and its active compounds. In some studies, taxonomic errors were encountered. Moreover, there is need for accurate methods in testing the safety and ethnomedicinal validity of Boerhavia species. Key words: B. diffusa, Boeravinones, Medicinal, Pharmacology, Punarnavine.

1

Abbreviations: IL-2 , Interleukin-2; NK, Natural Killer; TNF, Tumor Necrosis Factor; PBMCs, Peripheral Blood Mononuclear Cells; ROS, Reactive Oxygen Species; MAP, Mitogen-Activated Protein; ESR, Electron Spin Resonance; HBV, Hepatitis-B Virus; Th1, T helper 1; IFN, Interferon; WSSV, White Spot Syndrome Virus; VEGF, Vascular Endothelial Growth Factor; RT-PCR, Reverse Transcription Polymerase Chain Reaction; ELISA, Enzyme-Linked Immunosorbent Assay; HUVECs, Human Umbilical Vein Endothelial Cells; ATO, Arsenic Trioxide; NO, Nitric Oxide; IC 50, 50% Inhibition Concentration; EC50, Half Maximal Effective Concentration; ED50, Median Effective Dose; LC50, Half Maximal Lethal Concentration; HPLC-PAD, High-performance Liquid Chromatography with Pulsed Amperometric Detector; ESI-MS, Electrospray Ionization Mass Spectrometry; MMP, Matrix Metallo Peptidase; MOA, Monoamino Oxidase. EtOH, Ethanol; MeOH, Methanol; RSC50, Radical Scavenging Capacity of 50%; TDI, Toluene Diisocyanate; CMC, Carboxymethylcellulose. Chemical compounds studied it this review Boeravinone A (PubChem CID: 14018346) Boeravinone B (PubChem CID: 14018348) Boeravinone D (PubChem CID: 15081178) Boeravinone E (PubChem CID: 11537197) Boeravinone G (PubChem CID: 11537442) Boeravinone H (PubChem CID: 16745324) Coccineone B (PubChem CID: 44420939) Quercetin (PubChem CID: 5280343) Repenone (PubChem CID: 44257427) Repenol (PubChem CID: 44257428)

1. Introduction 1.1. Boerhavia genus: Boerhavia (frequently spelled ‘Boerhaavia’) is highly polymorphic genus of Nyctaginaceae also known as four-o’clock family because most of the species open their flowers four hours after noon i.e. in early evening or morning (Fosberg 1978; Levin et al., 2001). Nyctaginaceae encompasses 391 species in 32 genera. Due to taxonomic conflict and concepts, the genus Boerhavia includes variable number (20-40) of species (synonyms and homonyms) of subtropical or panatropical herbs (Fosberg 1978; Douglas and Manos, 2007). This has impacted scientific exploitations of some species of Boerhavia as only one species i.e. B. diffusa is predominantly studied. Most Boerhavia species 2

possess worldwide medicinal uses and hence occupied positions in different systems of medicine including Indian Ayurveda, Siddha and Unani, Martinican medicine, African medicine, traditional Chinese medicines as well as Indian and Brazilian pharmacopoeia. Six important species viz., B. diffusa, B. repens, B. chinensis, B. erecta, B. elegans (synonym: B. rubicund) and B. reniformis (synonym: B. rependa) are found in India (Chopra et al., 1969; Dev, 2006). Out of 180 isolated compounds from Boerhavia genus, B. diffusa shared about 131 compounds and for most of these compounds, it is currently an exclusive source. Following this 46 compounds have been isolated from B. erecta. The compounds from Boerhavia genus include characteristic chromoalkaloids, quinonolizidine alkaloids i.e. punarnavine, flavonoids, phenolic glycosides, phenolic acids, sterols and organic acids. Also many pharmacological activities in B. diffusa have been reported and there are some reviews which included information on B. diffusa. However, the present review also covers studies in other species of Boerhavia genus and recent pharmacological data of B. diffusa published in last 5-6 years. 2. Geographical distribution and botanical description Boerhavia species are widespread and the dispersal is mostly due to birds and human activity. The genus name Boerhavia was given in honour of Hermann Boerhaave, a famous Dutch physician of the 18th century. The distribution of Boerhavia species is in the warmer parts up to an altitude of 2000 m. Besides this, they are found in disturbed areas, waste places, roadsides, dry pinelands, among scrub on tropical reefs (Spellenberg, 2004). Although native to India and Brazil, B. diffusa is found in the tropical, subtropical and temperate regions of the world. This may imply that worldwide distribution of B. diffusa have helped for establishment of its broader ethnomedicinal spectrum and hence a material of interest for most industries and researchers. In India rakt punarnava (B. diffusa) is known to possess more medicinal importance than shweta punarnava (B. erecta). Alone in Kerala state of India, the demand of B. diffusa roots was 1, 150 metric tonnes in year 2000 and it is among the 46 medicinal plants sourced largely from wastelands (Tewari, 2000; Ved and Goraya, 2007; Pathak et al., 2012; Patil and Bhalsing, 2015).

However, considerable information on phytochemistry and

pharmacology of B. diffusa is available under the taxonomically inappropriate names such as Boerhavia diffusa Linn., Boerhavia Diffusa etc. All the names of Boerhavia species discussed in this review have been checked against www.theplantlist.org. Nevertheless, most scientific literatures provided different synonyms for Boerhavia species because of taxonomic errors or incorrectly indentified from local peoples. For example, in the literature there are many synonyms have been provided for B. diffusa such as Boerhavia glabrata, B. repens, B. erecta, B. rependa, B. procumbens etc. which are actually different species (Douglas and Manos, 2007; Selvaraj et al., 2012). Also, species name Boerhavia rependa Willd is used in literatures for which the accepted name is Boerhavia reniformis Chiov. In addition, studies are also conducted under the name Boerhavia chinensis (L.) Asch. & Schweinf. but the accepted species name is Boerhavia chinensis (L.) Rottb. 3

The species name Boerhavia paniculata has been recently used in literature for accepted name Boerhavia paludosa (Domin) Meikle. Beside the above taxonomic errors, the sources of error also seem to be those which are described by Rivera et al. (2014) and this can lead to an errorneous future research. Though Boerhavia is well polymorphic genus, a comprehensive and reliable taxanomic approache is still needed for identification of Boerhavia species. Few noteworthy studies using molecular tools such as ITS genes are recently carried out in this respect (Douglas and Manos, 2007; Selvaraj et al., 2012). The detailed geographical distributions of some traditionally used Boerhavia species are enlisted in Table 1. Table 1 List of selected plant species in Boerhavia Sr. No.

Name of the Species

Native Habitat/Origin and common name

Geographical distributions

References

Boerhavia chinensis (L.) Rottb. (Synonym: Boerhavia chinensis (L.) Asch. & Schweinf.) Boerhavia coccinea Mill. Boerhavia diffusa L.

Malaysia

All Old World tropics, including S. Malaysia and E. Java (distinctly restricted to regions subject to a seasonal climate)

Stemmerik, 1964

Brazil (pega pinto) India (Rakt Punarnava) and Brazil (Erva tostao)

Edeoga and Ikem (2002) Fosberg, 1978; Spellenberg, 2004

Boerhavia elegans Choisy (Synonym: B. rubicund) Boerhavia erecta L.

Iran (Sorhmard)

Brazil, Mexico, North America, Nigeria. (Worldwide distributed) Tropical, subtropical, temperate regions of the world including Mexico, American continent, Asia, Africa, Indian Ocean Islands, Pacific Islands, Australia, Egypt and Sudan. Africa, India, Pakistan, and Saudi Arabia

6.

Boerhavia hualienense S.H. Chen & M.J.Wu

Taiwan

Edeoga and Ikem, 2002; Chou et al., 2004 Chen et al., 2007

7.

Boerhavia glabrata Blume

Hawaii

8.

Boerhavia procumbens Banks ex Roxb.

Pakistan (Sentori)

(Widely distributed)America, Africa, Madagascar, India, Java, Malaysia, Philippines, China, Taiwan (Recently a new species) This species is endemic, occurring only in the east coast of Taiwan Java north to Micronesia, Ryukyu also to the Hawaiian Islands Pakistan, South West Asia and India

India (Punarnava)

Biligiri Rangana Hill ranges, Karnataka, India

Krishna and Shanthamma, 2004a

Indo-Pacific and

Asia, Africa and Mediterranean

Fosberg, 1978; Fadeyi

1.

2. 3.

4.

5.

9.

Boerhavia reniformis Chiov. (Synonym: Boerhavia rependa Willd.) 10. Boerhavia repens L.

Tropical America (erect spiderling)

Fosberg, 1978; Sadeghi et al., 2014;

Chen et al., 2007

Stemmerik, 1964; Singh et al., 1988 Abbasi et al., 2012;

4

11. Boerhavia paludosa (Domin) Meikle (Synonym: Boerhavia paniculata Rich.)

Africa (alena) Brazil

regions

et al., 1989

South America, Australia and Venezuela

Souza et al., 2014

3. Quality assessment Among many medicinally important member of Boerhavia, there seems to be continual dependence of herbal practitioners, researchers and industries on B. diffusa. However, in context of above discussed taxonomical errors and lack proper tools for taxonomical discrimition, B. diffusa often adulterated with related species like B. erecta, B. repanda, B. coccinea B. verticillata, B. repens, B. tetranda and B. albiflora. To address these problems, an attempt was made by Ferreres et al. (2005) through establishment of phenolic fingerprint i.e. chemical identity of B. diffusa using HPLC- PADESI/MS technique. They concluded that, geographical location or soil composition may vary the phenolic content of B. diffusa. However, recent approach suggested the use of DNA barcoding for quality assessment in B. diffusa due to its close morphological and phytochemical profile with some species. Plant nuclear rDNA regions like ITS1 (located between 5.8S rRNA and 18S rRNA genes), ITS2 (located between 5.8S rRNA and 25S rRNA genes) and chloroplast plastid gene psbA-trnH was used for phylogenetic analysis. Use of ITS regions (non-coding DNA) in plant taxonomy is related to its small size (600-700bp) and high rate of divergence. The study revealed that compared to ITS2 and psbA-trnH genes, ITS1 effectively distinguished B. diffusa from other 14 species of Boerhavia (Selvaraj et al., 2012). 4. Ethnomedicinal uses of Boerhavia Boerhavia encompasses very important species as evident from their broad spectrum of ethnomedicinal uses worldwide which are enlisted in Table 2. Out of these species, B. diffusa documented to possess most wide uses. Other members also have reported to possess important medicinal uses. B. diffusa has been documented in very old Indian Ayurvedic books of medicine such as the Charaka Samhita, Sushrita Samhita, Ashtaanga Hridaya and Chakradatta. These books have mentioned various benefits such as the book Chakradatta for treatment of chronic alcoholism, Bhaishajya ratnaavali for treatment of oedema and haematinic, Ashtaang Hridaya for stimulation of urinogenital systems etc. (Khare, 2004). In Uttarpradesh, India the local peoples used it as blood purifier, myocardial stimulant, expectorant, in jaundice, cough and snake bite (Singh et al., 2010). In Ayurveda and Unani, B. diffusa plant used to cure 22 different types of ailments. In Brazilian pharmacopeia, 23 traditional uses have been described for this plant, while in Africa and Middle East, the plant is prescribed for 14 ailments (Apu et al., 2012). In Martinican medicine, B. diffusa is a pain reducing agent and decoction or juice of leaves is used for its analgesic and anti-inflammatory properties (Robineau et al., 1996; Hiruma et al., 2000). The drug ‘punarnava’ include whole plant of 5

B. diffusa which is documented in Indian Pharmacopoeia as a diuretic (Chopra et al., 1969). It is also used in the treatment of anasarca, anaemia, scanty urine and ascites. The flowers and seeds are used as contraceptive (Chopra et al., 1956). Many commercial herbal formulations make use of B. diffusa plant for which the role or indications suggest its ethnomedicinal practises are valued (Table 3). B. chinesis roots are antihelmintic, cure leucorrhoea, use to ripen ulcers (Pullaiah 2006). In Southern Sudan, B. erecta root is used in treatment of remnant of newly detached umbilical cord in baby (Muzila, 2008). According to Baluch people, the plant of B. elegans is known to be a regenerator of heart and kidney and also, effectively used in urinary disorders, intestinal infections and diabetes (Ramazani et al., 2010). We have highlighted below important traditional uses reported for Boerhavia species which would be of potential interest for scientific study (Table 2).

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Table 2 Worldwide ethnomedicinal uses of important species of Boerhavia genus Name of the Species Boerhavia chinensis (L.) Rottb. (Synonym Boerhavia chinensis (L.) Asch. & Schweinf.)

Country/ Province

Boerhavia coccinea Mill.

Boerhavia diffusa L.

Ethnomedicinal Uses

Type of recipe

Reference

a) Antihelmintic

Oral administration of powdered root

Pullaiah, 2006

Northeast of a) Treatment of venereal disease, removing Brazil kidney\liver and urinary (Local system obstructions people)

infusion of the roots

Braga, 1960

West Africa a) Toothache

Inhalation of smoke from leaf pulp mixed with peanut oil Roots ground with other herbs and taken in water leafy twig

Muzila, 2008

Drinking tea made by boiling roots in water (sometimes interchanging with B. erecta) Infusion of stems with leaves.

Austin, 2010

Spongy roots decoction Decoction of Root

Bossard, 1996

Root decoction

Muzila, 2006

Nigeria

a) Vermifuge (to expel worms or other animal parasites from the intestines)

Cameroon (Local people) Mexico (Seri, Gurijiyo, Mayo peoples)

a) Pneumonia

Angola (regions of Cuanza Norte, Golungo Alto)

a) Used for arthritis, cramp, joint pain, rheumatism, kidney pain and as an antiinflammatory

Brazil (Local people)

Ghana

a) Measles

b) For cirrhosis, jaundice, hepatitis. a) Used as a diuretic, for urinary disorders like nephritis, abundant urine, albuminuria, gallstones, urinary retention, for hepatitis, jaundice, cystitis, ascites, Beriberi and blenorrhagia. a) Anaemia and

Jiofack et al., 2009

Bossard, 1996

Cruz, 1995

7

applied externally to yaws, b) Abdominal tumours

c) To treat heart troubles, palpitations and jaundice. India a) Asthma, scanty urine, and (Sahariya internal inflammation people) disorders, piles leucorrhoea, rheumatism, and stomach ache elephantiasis, to treat seminal weakness and blood pressure Nigeria a) Anticonvulsant, antiasthmatic, expectorant, and emetic. West Africa a) Sprain, blennorrage, (Ivory rheumatism, diffuse pains, Coast) maux of reins b) Asthma, Cholera, Paludism Iran

a) Gonorrhoea, nephritis and edema. b) Appetizer and to treat joint pain a) Gastro-enteritic problems,

Namibia (Bergdamar a people) (Damara b) Prolapsed uterus people) Congo a) Mumps, laryngitis and burns

Boerhavia elegans Choisy

Martinique

a) Analgesic and antiinflammatory

Iran

a) To treat dysmenorrhea, urinary tract disorders, intestinal infections, inflammation, jaundice, and body weakness in traditional medicine. Moreover, this species has expectorant, antidiabetic, motive and diuretic properties and is used as the regenerator of heart and kidney a) They use its decoction for removing fatigue and as an aphrodisiac and used for inflammation

Bamposht (East of Saravan) (Baluch peoples) Pakistan

a) Used in opthalmia, eye wounds and pain of the joints. b) Carminative and useful in

Powdered root with butter or oil. Decoction of root Root decoction

Mitra and Gupta, 1997

Decoction of leaves

Andesina, 1982

Decoction of roots

Bouquet, 1974

Decoction of leaves Whole plant decoction Infusion of leaves Chewing or boiled roots

Guessan, 2009 Zagari, 1992

Tea made from boiled root Root sap is rubbed on neck and throat Fresh juice or decoction of leaves

Muzila, 2008

Muzila, 2008

Muzila, 2008

Hiruma et al., 2000 Ramazani et al., 2010

Sadeghi et al., 2014. Zargari, 1987.

Leaves Fruits

Najam et al., 2008 Sandhu et al., 8

Boerhavia erecta L.

India (Malayali peoples)

muscular pain, lumbago scabies and hasten delivery. a) Asthma

India a) A diuretic to treat jaundice, enlarged spleen, gonorrhea (Local and other internal communitie inflammations. It is also used s) as stomachic, cardio tonic, hepatoprotective, laxative, antihelmintic (expels parasitic worms), febrifuge (reduces fever), and an expectorant. Soligas a) Infective hepatitis Peoples (Biligiri Rangana a) To ripen abscesses and Hill ranges, ulcers. Karnataka, India) Mali b) To treat liver problems, gastrointestinal diseases and infertility problems Niger a) Against fungal infections Benin Kenya

Tanzania

Boerhavia procumbens Banks ex Roxb.

a) To treat convulsions in children a) To treat diarrhea

a) To treat rheumatism and scabies

b) Conjunctivitis Burkina a) Antimalarial, antibacterial, Faso antiviral India (Gond a) Jaundice and Bhill peoples) b) Against edema, dropsy, and in dysmenorrhea. c) Used in the treatment of respiratory and pulmonary diseases, in disorders of liver, eyes, stomach, urinary, and throat. cleansing the bowel, reducing fevers, and for the killing of intestine helminthes d) To treat, anaemia, dropsy and gonorrhoea and also used as Eye tonic

2011 Smoke of plant powder

Anjalam et al., 2014

Root

Rameshkuma r and Ramakritinan, 2013

Roots with goat’s milk

Krishna and Shanthamma, 2004a Rameshkuma r and Ramakritinan, 2013 Muzila, 2008

Root paste is rubbed on the skin Leaves decoction Ash of the whole plant Whole plant decoction Aqueous extract of leaves Ash of the whole plant mixed with oil Sap of leaves Decoction of plant Decoction of fresh plant

Muzila, 2008

Leaves

Shah and Khan 2006 Qureshi et al., 2010

dried roots

Root decoction

Muzila, 2008 Muzila, 2008

Muzila, 2008

Muzila, 2008 Hilou et al., 2013 Lone et al., 2015

Katewa et al., 2004

9

Pakistan Boerhavia reniformis Chiov. (Synonym: Boerhavia rependa Willd.)

Boerhavia repens L.

e) Against swelling and Scorpion sting a) Jaundice and Hepatitis

India a) To cure jaundice Traditional practitioners (Yalandur, Chamarajan agar, Karnataka, India) India a) Jaundice (medicine practitioners )

b) Bronchial asthma

West Africa a) Asthma, Leprosy and Syphilis b) Ulcers, yaws c) Chickenpox

Central Africa Nigeria (Yoruba people)

a) Aphrodisiac or curing of Stomach ache b) To treat filarial infection a) (Ethnomedicinally considered effective and hence preferred over B. diffusa and B. erecta), Ecbolic, to cure jaundice, and poulticing sprains b) Albuminuria, asthma, depurative, diarrhoea, dropsy, epilepsy, erysipelas, hysteria, jaundice, measles, nerves and spasm

Root paste Whole plant Roots with butter milk

Katewa et al., 2004 Abbasi et al., 2009 Krishna and Shanthamma, 2004a

Crushed roots as well as a necklace of small pieces of roots are worn by patient Root juice mixed with chilli Decoction of leaves Roots Ground roots with seeds powder of Blighia sapida Root decoction

Wabale and Petkar, 2005

Root sap Decoction of leaves and whole plant pulp

Muzila, 2008 Muzila, 2008

Wabale and Petkar, 2005 Muzila, 2008 Muzila, 2008 Muzila, 2008

Muzila, 2008

Muzila, 2008

10

Table 3 Important commercially available herbal formulations in which B. diffusa is used Sr. No. 1.

Name of herbal/polyherbal formulation Himalaya Punarnava Capsules

Name of company

Role/ Indications

Himalaya

2. 3.

Diabecon Livgood Capsules

Himalaya LivgoodTM

4. 5. 6.

Punarnava Capsules, Organic Dabur Chyawanprash Junior 500G Immunowin

7.

Pain Nivaran Churna

8.

Femon C

9.

Hepatonej Syrup

10. 11.

Hridya (Heart Tonic) Herbal Tea Medohar (Anti-Obesity) Tea

Fushi Dabur Rajasthan Aushadhalaya Pvt. Ltd. Rajasthan Aushadhalaya Private Limited Ocean Herbal Pvt. Ltd. Nej Biotech, Nadiad, Gujarat Ayushkar Nirogam India Private Limited

Controls Swelling / Edema, Diuretic Used for diabetes Used in Liver diseases and as diuretic renewing and rejuvenating Supplement General debility and immunedeficiency Rheumatoid/Osteo Arthritis Treatments Antifibrinolytic, Hemostatic Jaundice, Hepatitis, Liver cirrhosis Cardioprotective tonic Prevents Obesity

5. Phytochemistry In view of ethnopharmacological uses and with the advancement of technologies like MS, LCMS, ESI-MS etc., many studies in Boerhavia species revealed number of important phytochemicals in different parts (Table 4). A general approach to isolate compounds includes extraction, partitioning in different solvents and bioactivity guided fractionation using silica gel chromatography.

11

Table 4 List of phytochemicals isolated from Boerhavia species Chemical Sr. category/cla No. ss/subclass 1. 1 Alkaloids:

2. Phenolics a) Flavonoids /Phenolic acids/glycosi des

Chemical compound

Punarnavine

Name of the Species

Source

Reference

B. diffusa

Root

B. repens

Root Root

Manu and Kuttan, 2009a Nandi and Chatterjee, 1974 Abbasi et al., 2012

2 3

Betanin Isobetanin

B. procumbens B. erecta B. erecta

4

Neobetanin

B. erecta

Stem bark

5

Eupalitin

B. diffusa

Whole plant

B. repens

Whole plant Whole plant Leaves, Stem, Flowers Whole plant

Li et al., 1996

Whole plant Whole plant

Maurya et al. 2007

B. repens

Whole plant

Li et al., 1996

B. diffusa

Whole plant

Maurya et al. 2007

B. diffusa

Whole plant

Maurya et al. 2007

B. diffusa


Maurya et al. 2007

6

Eupalitin-3-O-β-Dgalactopyranoside

B. diffusa B. diffusa

B. repens

7 8

9

10

11

12 13

Eupatilin-7-O-β-Dgalactopyranoside Eupatilin 7-O-α-Lrhamnopyranosyl (1→2) α-Lrhamnopyranosyl (1→6)- β-Dgalactopyranoside. Eupalitin 3-O-β-Dgalactopyranosyl-(1→2)-β-Dglucopyranoside Quercetin-3-O- β-Dglucopyranoside- 7-O- β-Dglucopyranoside Quercetin- 3-O-α-Lrhamnopyranosyl (1→6) - β-Dgalactopyranoside. 3,3ʹ,5-Trihydroxy-7methoxyflavone 3,4-dimethoxyphenyl1-O-β-D-glucopyranoside


B. diffusa

Stem bark Stem bark

Hilou et al., 2013 Stintzing et al., 2004 Stintzing et al., 2004 Maurya et al., 2007

Maurya et al. 2007 Mundkinajeddue et al., 2003 Li et al., 1996

Maurya et al. 2007

Maurya et al. 2007

12

14 15

16

17

18 19

4ʹ,7-dihydroxy-3ʹmethylflavone 3′,4′,5,7-tetrahydroxyflavone-3O-α-D-rhamnopyranosyl (1→6)O-β-D-glucopyranoside 4′,5,7-tetrahydroxy-3′methoxy flavones-3-O-α-Drhamnopyranosyl(1→6)O-β-Dglucopyranoside Kaempferol

6-methoxykaempferol 3-O-β-D(1→6)-robinoside Quercetin

20

Catechin

21 22 23

[(-)-epicatechin] Isorhamnetin Rutin

24

Myricetin

25 26 27 28 29

Narcissin Procyanidins Isoquercitrin isorhamnetin Isorhamnetin-3-O-β-Dglucopyranoside Isorhamnetin 3-O-α-l-rhamnopyranosyl-(1→ 6)-β-d-glucopyranoside Isovitexin Eupalitin 3-O-β-Dgalactopyranosyl-(1ʺʹ→ 2ʺ)-Oβ-D-galactopyranoside 5,7-dihydroxy-6-8-dimethoxy flavones (Borhavone) 3,4-Dimethoxyphenyl-1-O-β-Dapiofuranosyl-(1ʺʹ→3ʺ)-O-β-Dgalactopyranoside hexaacetate Quercetin 3-O-robinobioside

30

31 32

33 34

35 36 37 38

3,4-dihydroxy-5methoxycinnamoylrhamnoside Quercetin 3-O-(2″- rhamnosyl)robinobioside Kaempferol 3-O-(2″-

B. diffusa


Maurya et al. 2007

B. erecta

Whole plant

Nugraha, 2010

B. diffusa

B. procumbens B. erecta B. procumbens B. erecta B. erecta B. erecta B. procumbens B. procumbens B. erecta B. erecta B. erecta B. erecta B. erecta

Root , leaves Aerial parts Whole plant Whole plant Root , leaves Whole plant Leaves Whole plant Leaves Leaves Leaves Whole plant Whole plant Leaves Leaves Leaves Leaves Leaves

B. erecta

Aerial parts

Do et al., 2013a

B. erecta B. diffusa

Aerial parts Whole plant

Do et al., 2013a Maurya et al., 2007

B. diffusa

Roots

B. diffusa

Whole plant

Gupta and Ahmed, 1984 Maurya et al., 2007

B. diffusa

Leaves, roots Leaves, roots Leaves, roots Leaves,

B. erecta

B. erecta B. procumbens B. repens B. diffusa

B. diffusa B. diffusa B. diffusa

Nugraha, 2010

erreres et al , 2005 Do et al., 2013a Bokhari et al., 2015b Li et al., 1996 erreres et al , 2005 Bokhari et al., 2015b Petrus et al., 2012 Bokhari et al., 2015b Petrus et al., 2012 Petrus et al., 2012 Petrus et al., 2012 Bokhari et al., 2015b Bokhari et al., 2015b Petrus et al., 2012 Petrus et al., 2012 Petrus et al., 2012 Petrus et al., 2012 Petrus et al., 2012

erreres et al , 2005 erreres et al , 2005 erreres et al , 2005 erreres et al , 2005 13

41

rhamnosyl)-robinobioside 3,5,4′-trihydroxy-6,7dimethoxyflavone 3-O-galactosyl(1→2)glucoside [eupalitin 3-Ogalactosyl(1→2)glucoside] Kaempferol 3-O-robinobioside

42

Eupalitin 3-O-galactoside

B. diffusa

43

Punarnavoside

B. diffusa

Leaves, roots Leaves, roots Root

44

B. diffusa

Leaves

45 46 47

Phenol, 4, 6-di (1, 1dimethylethyl)-2methylAlkamide N-trans-feruloyltyramine Epicatechin

B. diffusa B. diffusa B. erecta

Root Root Stem

48

Quercetin diglycoside

B. erecta

Stem

49

Quercetin 3-O-rutinoside

B. erecta

Stem

50

Kaempferol diglycoside

B. erecta

Stem

51

Isorhamnetin diglycoside

B. erecta

Stem

52

Isorhamnetin 3-O-rutinoside

B. erecta

Stem

53

Isorhamnetin 3-O-glucoside

B. erecta

Stem

54

B. erecta

Stem bark

B. erecta

Stem bark

Nugraha et al., 2015

B. erecta

Stem bark

Nugraha et al., 2015

B. erecta B. erecta

Stem bark Stem bark

Nugraha et al., 2015 Nugraha et al., 2015

B. repens

65 66

Boerhaavic acid Boeravinone A

67

Boeravinone B

B. diffusa B. diffusa B. coccinea B. diffusa B. coccinea

Whole plant Leaves Leaves Leaves Leaves Leaves, roots Aerial parts Root Root Root Root

Nazir et al., 2011

60 61 62 63 64

3-methoxybenzoic acid 4-O-βglucoside 3-methoxyacetophenone 4-O-βglucopyranoside Isorhamnetin-3-O-rutinoside-7O-β-glucopyranoside Isorhamnetin-3-O-rutinoside 2,3-dihydroxypropyl-benzoate 3-O-β-[4ʺ-methoxy] Glucuronide 5,7,3'trihydroxycoumaronochromone Ferulic acid Syringic acid Gentisic acid O-coumaric acids Caffeoyltartaric acid

Stintzing et al., 2004 Stintzing et al., 2004 Stintzing et al., 2004 Stintzing et al., 2004 Nugraha et al., 2015

39 40

55 56 57 58

59

b) Rotenoids


B. diffusa

B. diffusa B. diffusa B. diffusa B. diffusa B. diffusa

roots Leaves, roots Leaves, roots

erreres et al , 2005 erreres et al , 2005 erreres et al , 2005 erreres et al , 2005 Jain and Khanna, 1989 Umamenaka et al., 2012 Do et al., 2013b Do et al., 2013b Stintzing et al., 2004 Stintzing et al., 2004 Stintzing et al., 2004

Tacchini et al., 2015 Daniel, 2006 Daniel, 2006 Daniel, 2006 erreres et al , 2005 Do et al., 2013b Kadota et al., 1988a Santos et al., 1998 Kadota et al., 1988a Santos et al., 1998 14

68

Boeravinone C

69 70 71

Boeravinone D Boeravinone E Boeravinone F

72

Boeravinone G

73 74

Boeravinone H Boeravinone I

B. diffusa B. erecta B. diffusa B. diffusa

Root Root Aerial parts Root Root Root Whole plant Root Aerial parts Root Root

75 76

Boeravinone K Boeravinone M

B. erecta B. diffusa

Aerial parts Root

77 78

Boeravinone N Boeravinone P

B. erecta B. erecta B. diffusa

Aerial parts Aerial parts Root

79

Boeravinone Q


Aerial parts Root

80

Boeravinone R

B. diffusa

Root

81

Boeravinone S

B. diffusa

Root

82

9-O-methyl-10hydroxycoccineone B 10-demethylboeravinone C

B. diffusa

Root

B. diffusa B. erecta B. diffusa B. diffusa B. diffusa

Root Aerial parts Root Root

Borelli et al., 2006 Do et al., 2013a Borelli et al., 2006 Borelli et al., 2006 Borelli et al., 2006

B. erecta B. erecta B. erecta

Aerial parts Aerial parts Aerial parts

Do et al., 2013a Do et al., 2013a Do et al., 2013a

Root Root Root Root Root Whole plant Whole plant Whole plant

Borelli et al., 2006 Messana et al., 1986 Ferrari et al., 1991 Ferrari et al., 1991 Santos et al., 1998 Ahmed et al., 1990

83

B. diffusa B. coccinea B. erecta B. diffusa B. diffusa B. diffusa B. repens

90

2ʹ-O-methylabroisoflavone 6-O-demethylberavinone H 9-O-methyl-10 hydroxycoccineone B Cucumegastigmane Kaempferol 3-O-rutinoside Quercetin 3-O-β-dglucopyranoside Coccineon B

91 92 93 94

Coccineon C Coccineon D Coccineon E Repenone

B. diffusa B. coccinea B. coccinea B. coccinea B. diffusa B. repens

95

Repenol

B. repens

96

Boerharotenoid B

B. repens

84 85 86 87 88 89

Kadota et al., 1988b Santos et al., 1998 Do et al., 2013a Borelli et al., 2006 Borelli et al., 2006 Borelli et al., 2006 Nazir et al., 2011 Borelli et al., 2006 Do et al., 2013a Borelli et al., 2006 Belkacem et al., 2007 Do et al., 2013a Bairwa et al. 2013a and Bairwa et al., 2013b Do et al., 2013a Do et al., 2013a Bairwa et al. 2013a and Bairwa et al., 2013b Do et al., 2013b Bairwa et al. 2013a and Bairwa et al., 2013b Bairwa et al. 2013a and Bairwa et al., 2013b Bairwa et al. 2013a and Bairwa et al., 2013b Borelli et al., 2006

Ahmed et al., 1990 Nazir et al., 2011 15

d)Coumaron ochromonoid s

3.Terpenes

97

Boeravinone O

B. erecta

Aerial parts

Do et al., 2013a

98

Boeravinone J

B. diffusa

Root

99 Boeravinone L 100 Coccineon A 101 Boerharotenoid A

B. erecta B. erecta B. coccinea B. repens

102 Boerhavisterol 103 Camphor


104 Isomenthone

B. diffusa

105 Limonene

B. diffusa

106 Menthol

B. diffusa

107 Phellandrene

B. diffusa

108 Safranal

B. diffusa

109 α-Pinene

B. diffusa

110 Geranylacetone

B. diffusa

111 cis 4-Hexen-1-ol

B. diffusa

112 trans 2-Octanalc

B. diffusa

113 2-Nonen-1-olc

B. diffusa

114 2-Decen-1-ol

B. diffusa

115 Methylpyrrole

B. diffusa

116 3-Phenyl-2-(20 -pyridyl)-indole 117 Indole

B. diffusa

118 Eugenol

B. diffusa

119 β-Cyclocitral

B. diffusa

120 β-Ionone

B. diffusa

121 Dihydroactinidiolide

B. diffusa

122 Stigmasterol 123 Campesterol 124 β-sitosterol


Aerial parts Aerial parts Root Whole plant Roots Roots, leaves Roots, leaves Roots, leaves Roots, leaves Roots, leaves Roots, leaves Roots, leaves Roots, leaves Roots, leaves Roots, leaves Roots, leaves Roots, leaves Roots, leaves Roots, leaves Roots, leaves Roots, leaves Roots, leaves Roots, leaves Roots, leaves Root Root Root

Belkacem et al., 2007 Do et al., 2013a Do et al., 2013a Messana et al., 1986 Nazir et al., 2011

B. diffusa

B. erecta

Whole plant

Gupta and Ali, 1998 Pereira et al., 2009 Pereira et al., 2009 Pereira et al., 2009 Pereira et al., 2009 Pereira et al., 2009 Pereira et al., 2009 Pereira et al., 2009 Pereira et al., 2009 Pereira et al., 2009 Pereira et al., 2009 Pereira et al., 2009 Pereira et al., 2009 Pereira et al., 2009 Pereira et al., 2009 Pereira et al., 2009 Pereira et al., 2009 Pereira et al., 2009 Pereira et al., 2009 Pereira et al., 2009 Kadota et al., 1989 Kadota et al., 1989 Misra and Tiwari, 1971 Nugraha, 2010 16

B. repens

4. Saponin 5. Lignans

6. Others

Whole plant Root Root

Ahmed et al., 1990

125 α-2-sitosterols 126 Ursolic acid


127 β-amyrin

B. diffusa

128 β-amyrin acetate

B. diffusa

129 3-acetoxy-α-amyrin 130 4, 10-dihydroxy-8methoxyguai-7(11)-en-8,12olide 131 Oleanolic acid heteroside 132 Liriodendrin 133 Syringaresinol mono-β-Dglucose 134 Myo-lnositol,4-C-methyl-1,14Tetradecanediol 135 1-pentadecyne


Whole plant Whole plant Aerial parts Aerial parts


Roots Roots Roots

Bep, 1986 Lami et al., 1992 Lami et al., 1992

B. diffusa

Leaves

B. diffusa

Leaves

136 Phytol

B. diffusa

Leaves

B.repens

Umamenaka et al., 2012 Umamenaka et al., 2012 Umamenaka et al., 2012 Ahmed et al., 1990

137 3,5-Bis(trimethylsilyl)-2,4,6cycloheptatrien-1-one 138 Androstane-11, 17-dione, 3[(trimethylisilyl)oxy]-17-[O(phenylmethyl)oxime], (3a,5a) 139 hypoxanthine 9-L-arabinose

B. diffusa

Whole plant Leaves

B. diffusa

Leaves

B. diffusa

Roots

140 myricyl alcohol

B. diffusa

Roots

141 myristic acid

B. diffusa

Roots

142 β-ecdysone 143 Anthocyanins

B. diffusa B. erecta

Root Stem bark

144 Squalene

B. repens

145 Procyanidin B1

B. erecta

Whole plant Stem

146 Procyanidin B2

B. erecta

Stem

147 Dimeric procyanidin

B. erecta

Stem

148 149 150 151

B. diffusa B. diffusa B. diffusa B. diffusa

Roots Roots Roots Roots

B. diffusa

Roots

Boerhadiffusene Diffusarotenoid boerhavilanastenyl benzoate Boerhavine

152 Borhavone

Ranjini et al., 2013 Misra and Tiwari, 1971 Maurya et al. 2007 Maurya et al. 2007 Do et al., 2013b Do et al., 2013b

Umamenaka et al., 2012 Umamenaka et al., 2012 Ojewole and Andesina, 1985 Ojewole and Andesina, 1985 Ojewole and Andesina, 1985 Suri et al. 1982 Marulkar et al., 2012 Ahmed et al., 1990 Stintzing et al., 2004 Stintzing et al., 2004 Stintzing et al., 2004 Gupta and Ali, 1998 Gupta and Ali, 1998 Gupta and Ali, 1998 Ahmed and Yu, 1992 Gupta and Ahmed, 1984; Ahmed and Yu, 1992 17

153 154 155 156

B. diffusa B. diffusa B. diffusa B. diffusa

Roots Aerial parts Aerial parts Aerial parts

Tacchini et al., 2015 Do et al., 2013b Do et al., 2013b Do et al., 2013b


Aerial parts Aerial parts Aerial parts

Do et al., 2013b Do et al., 2013b Do et al., 2013b

B. diffusa

Aerial parts

Do et al., 2013b

B. diffusa

Aerial parts

Do et al., 2013b

B. diffusa

Aerial parts

Do et al., 2013b

B. diffusa

Aerial parts

Do et al., 2013b

172 Hentriacontane

B. diffusa B. diffusa B. diffusa B. diffusa B. diffusa B. diffusa B. diffusa B. procumbens B. diffusa

Roots Roots Roots Roots Roots Roots Roots Whole plant Roots

173 Triacontanol 174 Quinic acid


175 Fumaric acid

B. diffusa

176 Ketoglutaric acid

B. diffusa

177 Pyruvic acid

B. diffusa

178 Oxalic acid

B. diffusa

179 Vitamin E acetate

B. diffusa

Roots Roots, leaves Roots, leaves Roots, leaves Roots, leaves Roots, leaves Leaves

Kadota et al., 1989 Kadota et al., 1989 Kadota et al., 1989 Kadota et al., 1989 Kadota et al., 1989 Kadota et al., 1989 Kadota et al., 1989 Bokhari et al., 2015b Misra and Tiwari 1971 Suri et al., 1982 Pereira et al., 2009

157 158 159 160 161 162 163 164 165 166 167 168 169 170 171

Vanillin Allantoin Sophorophenolone N-trans-feruloyl-3Methyldopamine (+)-zedoalactone A Ciwujiatone 1-β-D-glucopyranosyloxy-3,5dimethoxy-4-hydroxybenzene 1-β-D-glucopyranosyloxy-3,4dimethoxybenzene 1-β-D-glucopyranosyloxy-1phenylmethane 1-β-D-glucopyranosyloxy-2pheny-lethane 1- β-D-glucopyranosyloxy-2methoxy-4-ethanoylbenzene Palmitic acid Heptadecyclic acid Oleic acid Strearic acid Arachidic acid Behenic acids Sterol esters Caeffic acid

Pereira et al., 2009 Pereira et al., 2009 Pereira et al., 2009 Pereira et al., 2009

Umamenaka et al., 2012 180 Potassium nitrate B. diffusa Leaves Kokate et al., 2005 Note: the serial number given in the table corresponds to the number given (bold) in brackets in the discussion part. 5.1. Alkaloids The phytochemical studies in Boerhavia genus seems to be carried out because of its immense use in traditional medicines across the world. Therefore, it is obvious to search for alkaloids (1-4) in this genus as alkaloids are well known to possess important biological activities. The characteristic chromoalkaloids known as betacyanins are located in the cellular vacuoles of subepidermal tissues hence the stem barks of Boerhavia species appear red (Stemmerik, 1964). Evaluation of antioxidant 18

potential in B. erecta showed that these betacyanins i.e. betanins (2-4) were most potent antioxidants (Stintzing et al., 2004; Hilou et al., 2013). This illustrates why traditionally the redder species are more preferable in disease treatments. Total quantity of betacyanin alkaloids reported from B. erecta was 186 mg/100 g (Stintzing et al., 2004). However, the most promising bioactive alkaloid found in Boerhavia genus is a type of quinolizidine alkaloid known as punarnavine (1) having chemical formula C17H22N2O and M.P. of 236°–237°C (Agarwal and Dutt, 1935; Manu and Kuttan, 2009a; Manu and Kuttan, 2009b; Saraswati et al., 2013; Dhingra et al., 2014). Punarnavine has been crystallized but its molecular structure is yet to be elucidated because most studies rely on old protocol (Agarwal and Dutt, 1935) and require advanced techniques. Maximum punarnavine accumulation was found in different parts of the B. repens as the plant gets older (Nandi and Chaterjee, 1974). In another study total alkaloid content (estimated as punarnavine content) in the root of three year old mature plant of B. diffusa was higher (2%) than in vitro regenerated roots (0.15%) (Shrivastava et al., 1995). Many factors such as stress, endogenous cytokinins are known to promote alkaloids which come into play as the plant ages and suggested to have physiological or regulatory role in metabolism of plants. Recently, ethanolic extract of whole plant of B. diffusa provided total alkaloid content 0.232 gm per 100gm (0.2%) (Beegum et al., 2014) while same extract contain punarnavine as 0.01% (Manu and Kuttan, 2009a).

Betanin PubChem CID: 54600918 Fig.1. Alkaloid from Boerhavia diffusa 5.2. Phenolic compounds: Phenolic acids/glycosides, Flavonoids and Rotenoids Although alkaloids were early detected in an approach to assess ethnomedicinal values of Boerhavia species recent studies indicate that flavonoids (including rotenoids) are responsible for most of these traditional uses. The total flavonoid content in B. diffusa was found to be 5.651 g/100 g (5.6%) which is many folds higher than alkaloids (0.2%) (Beegam et al., 2014). Approximately 97 (5-101) phenolic compounds which includes flavonoids, rotenoids, phenolic acids and phenol glycosides (Table 4) attributing many different biological activities are isolated from Boerhavia species. Traditional use of 19

Boerhavia species as a laxative is related to its high content of phenolic compounds (Fadeyi et al., 1989). Phenolic content can also be correlated with antioxidant property of plant species and presence of tannins contributes to astringency (Ma et al., 2014). B. diffusa contains moderate tannins (16 mg/gm) and high phenols (2.471 g/100 g) (Beegam et al., 2014). Punarnavoside, a phenolic glycoside isolated from B. diffusa act as an antifibrolytic agent. Phenolic compounds such as ferulic acid and vanillin have been reported with important biological activities in B. diffusa (Tacchini et al., 2015). Biactivity guided fractionation led to the isolation of eupalitin-3-O-β-D-galactopyranoside from B. repens (14.4 mg/1.2 kg) (Li et al., 1996). Another study claimed that for large scale isolation of eupalitin-3-O-β-D-galactopyranoside, B. diffusa sample from Jammu region, India (yield = 3g/1kg) can be more promising than that of B. repens (Mundkinajeddue et al., 2003). From this study, it can be inferred that geographical location of B. diffusa may affect its phytochemical productivity. A partition technique using different solvents was carried out for enrichment of flavonoids from methanolic extract of B. diffusa roots (Mahesh et al., 2011). Recently, spectroscopic analysis of Boerhavia erecta stem bark has proven to be a potential source for bioactive molecules (54-58). (Nugraha et al., 2015).

Name

R1

R2

R3

R4

R5

R6

Eupalitin

H

OH

H

OH

OCH3

CH3

Eupalitin-3-O-β-D-galactopyranoside

H

OH

H

a

OCH3

CH3

Quercetin (PubChem CID: 5280343)

H

OH

H

H

H

H

Isorhamnetin (PubChem CID: 5281654)

CH3

OH

H

H

H

H

Isoquercetin (PubChem CID: 5280804)

H

OH

H

b

H

H

20

Isorhamnetin-3-O-β-D-glucopyranoside

OCH3

OH

H

b

H

H

H

OH

OH

b

OCH3

b

(PubChem CID: 5318645) Quercetin-3,7-di-O-glucoside 3,3',5-trihydroxy-7-methoxyflavone

OH

H

H

OH

H

CH3

Eupalitin-7-O-α-rhamosyl(1→2)α-

H

OCH

OCH

H

OCH3

c

3

3

OH

OH

d

H

H

OH

H

d

H

H

rhamnosyl(1→6)-β-D-galactopyranoside Quercentin-3-O-α-L-

H

rhamnopyranosyl(1→6)-β-Dgalactopyranoside Rutin (PubChem CID: 5280805)

H

Narcissin (PubChem CID: 5481663)

OCH3

OH

H

d

H

H

Myricetin (PubChem CID: 5281672)

OH

OH

OH

H

H

H

OH

H

H

H

H

Kaempferol (PubChem CID: 5483905)

H

Name 4',7-dihydroxy-3'-methylflavone Isovitexin

R1

R2

R3

CH3

H

H

OH

b

(PubChem H

CID:162350)

Boerhaavic acid

3,4-dimethoxyphenyl-1-O-β-D-apiofuranosyl-(1''→3')-O-β-D-glucopyranoside

21

Punarnavoside Fig. 2. Chemical structures of flavonoids from Boerhavia genus 5.2.1. Rotenoids Rotenoids are a group of isoflavonoid type of compounds less acutely toxic in mammals and can be act as bioinsecticide (Wagner et al., 2012). Rotenoids seem to be of interest for elaborative research in this genus as they accounted for most of the pharmacology or ethnomedicinal claims of Boerhavia species (Borelli et al., 2006; Belkacem et al., 2007; Aviello et al., 2011; Bairwa et al., 2013a; Bairwa et al., 2013b; Do et al., 2013a; Do et al., 2013b). Using Kupchan partitioning and bioactivity guided fractionation technique 30 rotenoids (66-96) and 5 coumaronochromonoids (96-100) have been isolated from three different plants viz., B. diffusa, B. erecta and B. repens (Borelli et al., 2006; Belkacem et al., 2007; Bairwa et al., 2013a; Bairwa et al., 2013b; Do et al., 2013a; Do et al., 2013b ) (Table 4, Fig. 5). Rotenoids can be linked to isoflavonoids as biosynthesis of isoflavones involves a characteristic step of migration of phenyl from the 6a carbon atom in rotenone to 12a carbon atom. So, C-6 is not inherent but an additional carbon in rotenoids possibly supplied by methionine (Crombie, 1984). All rotenoids possess rotoxen skeleton (Fig. 3. i.) and the best known naturally occurring rotenoid is rotenone. Rotenone is mostly used as insecticide and fish poison but can act as anticancer agents possibly by inhibiting mitochondrial oxidation of NADH (Crombie, 1984; Crombie et al., 1992; Belkacem et al., 2007; Fig. 4). However, rotenoids in Nyctaginaceae differes from that occur in Leguminosae in which former lack isoprenoid residue on ring D, possess either mono- or no substitution on ring A and sometimes methoxy substituted at C-10. Loss of C-6 in rotenoids because of bioenegetically direct cyclization of isoflavonoids leads to coumaronochromonoids such as boeravinone J, boeravinone L, boeravinone O and coccineone A. All these structural features in rotenoids from Nyctaginaceae are unfavourable for their cytotoxic activity (Fig. 4) (Belkacem et al., 2007; Kadota et al., 2007; Do et al., 2013a and Do et al., 2013b). On the other hand, boeravinone G (72) which is methoxy substituted at C-9 rather than C-10 showed most potent anticancer and antioxidant activities (Belkacem et al., 2007; Aviello et al., 2011). Among five new rotenoids (boeravinone M, P, Q, R, S) from B. diffusa, boeravinone S (81) was observed to be most potent in vitro COX-1 and COX-2 inhbitory agent while significant in vivo anti-inflammatory potential observed for boeravinone B (67) (Bairwa et al., 2013a; .Bairwa et al., 2013b). Microwave-assisted extraction (MAE) technique (a rapid method over conventional soxhlet and maceration) has estimated 22

yields of boeravinone B (0.15%) and boeravinone E (70) (0.32%) in B. diffusa (Bhope et al., 2011). However, advanced study in B. diffusa using RP-HPLC has estimated 0.22% and 0.05% yields of boeravinone B and boeravinone E respectively (Bhope et al., 2013). Later study was supported by Bairwa et al. (2014) in which boeravinone B was detected as a major compound when quantified by UPLC/PDA technique in three different geographically located samples of B. diffusa.

i.

ii.

Fig. 3. i. Rotexon skeleton ii. General rotenoid structure of Boerhavia spp.

Fig. 4. General structural features of i. Rotenone from Leguminosae and Fabaceae ii. Rotenoid from Boerhavia spp. (Nyctaginaceae) iii. Coumaronochromonoid from Boerhavia spp. shown positivity (pink) and negativity (green) in cytotoxic activity.

Name

R1

R2

R3

R4

R5

R6

Boeravinone A

H

H

CH3

H

CH3

OCH3

H

H

H

H

CH3

CH3

(PubChem CID: 14018346) Boeravinone B

23

(PubChem CID: 14018348) Boeravinone D

OH

H

CH3

H

H

CH3

OH

H

H

H

H

CH3

H

OH

CH3

H

CH3

H

H

OH

CH3

H

CH3

CH3

H

H

H

OH

H

CH3

Boeravinone M

H

OH

H

H

CH3

H

Boeravinone P

H

H

CH3

H

H

H

Boeravinone Q

H

H

CH3

OCH3 H

CH3

Boeravinone R

H

H

H

OCH3 H

CH3

Boeravinone S

OH

H

H

H

H

H

Coccineone B (PubChem CID:

H

H

H

H

OH

H

6-O-demethylboeravinone H

H

OH

H

H

CH3

CH3

Repenone (PubChem CID: 44257427)

H

H

e

H

H

OH

Repenol (PubChem CID: 44257428)

OH

H

e

H

H

OH

Boerharotenoid B

H

OH

H

H

H

H

5,7,3'-trihydroxycoumaronochromone

H

H

H

H

OH

CH3

(PubChem CID: 15081178 ) Boeravinone E (PubChem CID: 11537197) Boeravinone G (PubChem CID: 11537442) Boeravinone H (PubChem CID: 16745324) Boeravinone I (PubChem CID: 16203334)

44420939)

Name

R1

R2

Boeravinone C

OH H

R3

R4

CH3 OH

(PubChem CID:13940642) Boeravinone K

H

CH3 CH3 OH

10-demethylboeravinone C

OH H

Boeravinone N

OH CH3 H

Coccineone E

H

CH3 OH b

CH3 CH3 OH

(PubChem CID: 12004176) Coccineone C

OH CH3 H

OH

Coccineone D

OH H

H

OH

R2

R3

Name

R1

24

Boeravinone J (PubChem CID:

OH

H

CH3

Boeravinone L

OH

H

H

Boeravinone O

a

H

H

Coccineone A

H

OH H

Boerharotenoid A

H

H

12004175)

Boeravinone F

CH3

Diffusarotenoid

Fig. 5. Chemical structure of rotenoids from Boerhavia genus

5.5 Terpenes A ‘flavour fingerprint’ of plant species generally recognized by animals and humans can be derived from a modified form of terpens so called ‘terpenoids’ (Pereira et al., 2009). The term ‘terpenes’ and ‘terpenoids’ are derived from ‘turpentine’. The five carbon units in terpenoids are called isoprene units (give off the gas isoprene at high temperatures) or

hemiterpenes as C-10 terpenoids

(Monoterpenes) were initially thought to be smallest group of this class (Buchanan et al., 2015). Terpenes are components of essential oil and possess important biological activities. In view of this,

25

approximately 28 terpenes (102-130) have been reported from leaves and roots of B. diffusa (Pereira et al., 2009). Structures of some important terpenes have been shown in Fig. 6.

R = H (β-sitosterol) R = Glucose (β-sitosterol-β-D-glucoside)

4,10-dihydroxy-8-methoxyguai-7(11)-en-8,12-olide

Ursolic acid Fig. 6. Chemical structure of terpenes from Boerhavia genus

5.6 Lignans Liriodendrin (132) and Syringaresinol mono-β-D-glucose (133) were the two major lignans detected from B. diffusa (Lami et al., 1992). Recent approach has evaluated the bioactive potential of liriodendrin isolated from B. diffusa using PASS (Prediction of Activity Spectra for Substances) assessment technique. It was found that out of total 22 reported activities of B. diffusa, 10 activities including smooth muscle relaxant, reproductive dysfunction, vasoprotector and emetic properties were predicted by PASS for liriodendrin with prediction coefficient of 0.45. The remaining activities were predicted by PASS for other compounds of this plant gaining the overall coefficient of 0.77 (Goel et al 2011).

26

Name of the Compound

R1

R2

Liriodendrin

b

b

Syringaresinol mono-β-D-glucoside

b

H

Fig. 7. Chemical structure of lignans from Boerhavia genus 5.7. Saponins Although different saponins were detected in other Nyctaginaceae members there are meagre studies in saponins from Boerhavia. One current study estimated moderate amount of saponins (1.59%) in the chloroform extract of whole plant of B. diffusa (Beegam et al., 2014). Only one triterpenoid saponin i.e. oleanolic acid heteroside was reported from B. diffusa which showed anti-inflammatory activities (Bep, 1986). 6. Pharmacological Properties Although substantial pharmacological data in the literatures is available for Boerhavia genus, many pharmaceuticals activities essentially lack appropriate comparisons with positive controls. Also, the studies carried out were inappropriate with relevant pharmacological doses of active extracts as well as positive controls and their time duration, source of materials used and minimum effective dose. Due to its wide medicinal uses in traditional systems the B. diffusa has been executed from long for its ethnopharmacological potential and many reviews included this information. However, here (Table 5) we have enlisted available information on Boerhavia species and only discussed below (Section 6.1) the recent data published in last 5-6 years. Table 5 Pharmacological activities reported for Boerhavia species

Species

Pharmacol ogical Activity

Type

Assayin g parame ters

Boerhav ia

Antimalari al

In vitro

parasite lactate

Par ts use d Aeri al

Dose range

Results/Outcomes

Bioactive metabolite and/type of extract 4.68 In vitro anti-plasmodial Dichlorom to 50 activity observed with ethane

Referen ces

Ramaza ni et al., 27

elegans

Boerhav ia erecta

and dehydro part in vivo genase (pLDH) assay and 4day suppress ive test in mice

Antioxidan t

In vitro

DPPH, FRAP

Lea ves

Cytotoxicit y

In vitro

96-Well cell cytotoxi city assay

Lea ves

Antimalari al

In vivo

Blood schizont icidal activity

Ste m bark

Heptoprote ctive

In vivo

Roo ts

Antioxidan t

In vitro

Serum enzyme such as ALT, AST and ASP SOD, catalase and peroxida se

Lea ves

μg/ml

IC50 values 15.33 ± 0.07 µg/ml and 11.97 ± 0.05 against two strains of Plasmodium falciparum K1 (chloroquineresistant strain) and CY27 (chloroquine-sensitive strain) using pLDH assay and also in vivo assay in mice inoculated with Plasmodium berghei (ANKA strain) showed anti-plasmodial activity with IC50 values of < 20 μg/ml Significant antioxidant activity with IC50 = 6.85 μg/ml

50 μg/ml

extract partitioned from ethanol extract

Phenolics in MeOH extract Phenolics in MeOH extract

Very low cytotoxicity against MCF-7 at 50 μg/ml of extract compared to standard drug Doxorubicin (10μg/ml) 50Red stem bark of B. Decoction 1000 erecta against mg/kg Plasmodium berghei /day berghei in mice showed low acute toxicity (ED50 value of 564.95±6.23 mg/kg and LD50 value of 2148mg/kg) compared to standard drug chloroquine (ED50 value of 14.59±3.2 mg/kg) when observed on 4th day. 100m Levels of serum bilirubin, 50% EtOH g/100 total protein, albumin and extract g b.w. also serum enzymes were for 25 restored in CCl4 days hepatotoxic male Wistar rats (175-200 gm weight). B. erecta exhibited enzymic antioxidant activities such as superoxide dismutase (SOD) catalase (CAT)

2010

Sadeghi et al., 2014. Sadeghi et al., 2014.

Hilou et al., 2006

Krishna and Shantha mma, 2004a

Phenolics Rajeswa and or ri et al., flavonoids 2010 in MeOH extract 28

activity

Boerhav ia procumb ens

In vitro

ABTS

ste m bark

Antihelmin tic

In vitro

Paralysi Roo s time t and death time

Anticancer

In vitro

antiproli Aeri ferative al Sulforho part damine B (SRB) assay

Antioxidan t

In vitro

Antiinflammato ry

In vivo

DPPH, ABTS, superoxi de, OH, H2O2 and phospho molybda te radical scavengi ng activity carragee nan induced Paw

Wh ole plan t

Wh ole plan t

300, 250, 200, 150, 100, 50, and 25 µg/ml

and peroxidase (POD) 0.086 ± 0.009, 0.440 ± 0.350, 40.95 ± 12.60 units/mg protein respectively 70% MeOH extract showed higher antioxidant activity of betacyanin fractions than phenolic compounds (RSC50 = 171.2 ± 8.4) At 20 mg/g extract showed in vitro antihelmintic activity against Pheretima posthuma with paralysis time of 3.25±0.21 min. and death time of 19.03±0.25 min. Rotenoids purified from ethyl acetate extract were found to exhibit significant cytotoxicity against HeLa (human epithelial carcinoma) and MCF-7 (human breast cancer) cell lines at the concentration of 100 μg/ml with camptothecin as the positive control. Various assays showed antioxidant potential

Betanin

Hilou et al., 2013

Tannins in Marulka MeOH r et al., extract 2011

Boeravinon e C, K, (HeLa cells) Boeravinon e M (MCF7 cell line) 10methylboer avinone C (both) Polyphenol s in nbutanol fraction

Anti-edemous effect was MeOH observed at all stages of extract edema development which led to the

Do et al., 2013a

Bokhari et al., 2015b

Bokhari et al., 2015b

29

edema.

Boerhav ia rependa

Hepatoprot ective

In vivo

Serum Roo enzyme ts such as ALT, AST and ASP

In vivo

Serum enzymes

Lea ves and call us

conclusion that both early and delayed phases of carrageenan-induced inflammation were attenuated may because of histamine or NO release Levels of serum bilirubin, total protein, albumin and also serum enzymes were restored in CCl4 hepatotoxic male Wistar rats although B. erecta roots extract was found to be more effective than B. rependa Levels of serum enzymes, serum albumin etc., were restored in Wistar rats and evaluated that leaf calli can be efficiently used as alternative to in vivo leaves for hepatoprotective purpose

50% EtOH Krishna extract and Shantha mma, 2004a

EtOH extract

Krishna and Shantha mma, 2004b

6.1. Recent Pharmacological Investigations in Boerhavia 6.1.1. Effect on metabolism 6.1.1.1 Antidiabetic activity The global prevalence of diabetes is increasing and can lead to secondary complications such as blindness, increased risk for cardiovascular diseases and also kidney failure. Moreover, 80% deaths occur in middle- and low-income countries due to diabetes (WHO 2015). Most of the peoples in these countries mainly rely on traditional or indigenous medicines, especially in ailments such as diabetes for which Western medicines are associated with severe side effects (Zuang et al., 2013). In such circumstances, it is utmost important to evaluate the scientific validity and effectiveness of these traditional medicines. Oral admistration of Boerhavia diffusa (at high dose of 400 mg/kg b.w.) reduced the blood glucose concentration when observed on 7th, 14th and 21st day compared to negative control (received 0.5 ml of 5% Tween 80) diabetic rats. To explain a high dose of Boerhavia extract for antidiabetic assay, they also carried out the toxicity studies at this high dose and said that at a dose of 400 mg/kg b.w. B. diffusa is safer to use. The positive control rats in this study received

30

glibenclamide (0.5 mg/kg b.w.). According to one study, methanolic extract of B. diffusa is more effective than ethanolic extract (each at a dose of 200 mg/kg b.w.) in regenerating damaged β-cells of pancreas in streptozotocin as well as alloxan treated male diabetic rats. This study used glibenclamide (10 mg/kg b.w.) as positive control (Bhatia et al., 2011). A study of aqueous extract of B. diffusa in alloxan-induced diabetic rats observed 38.07%, 51.95% and 21.56% reduction of glucose level respectively at the doses of 100, 200 and 400 mg/kg in 6 hours (Chude et al., 2011). They postulated that this hypoglycaemic activity was due to pheripheral utilization of glucose. However, the study requires the positive controls for comparative efficacy. 6.1.2. Effect on immune system: immunomodulation 6.1.2.1. Immunosuppressive activity Immunomodulation is the regulation of immune responses by stimulating them to prevent infectious diseases or by suppressing them in the undesired conditions. Boerhavia genus has long been used traditionally in treatment of ulcers and asthmic conditions but require scientific validation. In recent study, 95% ethanol extract of B. diffusa has shown to suppress inflammatory response both in vivo (mouse gastritis ulcer model) and in vitro (LPS-treated RAW264.7 cells) in a dose dependent manner (20 to 200mg/kg b.w.) which is comparable to standard drug ranitidine (40 mg/kg b.w.). The study further evaluated molecular inhibitory mechanisms lying behind this immunosuppressive activity and it was found that B. diffusa extract and its active ingredient luteolin markedly suppressed the activation of NF-κB by blocking a series of signalling cascades ranging from Syk/Src to IκBα The AP-1 signalling pathway, which is controlled by TRAF6/TAK1, was also inhibited by the extract (Thai et al., 2015). Also, methanolic extract of B. procumbens at a dose of 200mg/kg b.w. showed anti-asthmatic activity in TDI provoked rats by inhibiting infiltration of eosinophils and lymphocytes in lungs. Dexamethasone (2.5 mg/kg b.w.), a steroid medication was used as positive control in this study (Bokhari and Khan, 2015). These results support the ethnomedicinal practises of Boerhavia genus in immune reactions such as allergies, asthma, inflammation and ulcers. 6.1.2.2. Immunostimulatory activity Interestingly, the alkaloidal fraction of B. diffusa showed immunostimulatory activity in Balb/c mice. This alkaloid fraction possessed ‘punarnavine’ which acquired its name from punarnava (a Hindi name for Boerhavia diffusa) plant. Punarnavine dose (40 mg/kg b.w.) in Balb/c mice for 5 days inhibited the production of proinflammatory cytokines such as TNF-α, IL-1β and IL-6. It has shown to stimulate immune system with enhanced stem cell proliferation, stem cell differentiation and antibody formation. In this study, the results were compared with untreated control mice (Manu and Kuttan, 2009b).

31

6.1.3. Effect on renal disorders: 6.1.3.1. Nephroprotective effect The studies in mercury chloride toxicity rats demonstrated that 200 mg/kg b.w. aqueous leaf extract of B. diffusa was given orally for 5 days effectively protect kidneys from damage (Indhumathi et al., 2011). In eupalitin-3-O-β-D-galactopyranoside treated Koi carp (Cyprinus carpio) fish, the levels of urea, creatinine and marker enzymes were normal and no pathological changes in renal tissue were observed (Fathima et al., 2012). Although these studies lack of positive controls, their results suggested the traditional significane of Boerhavia diffusa in urinary disorders. A recent study in gentamicin-induced nephrotoxicity rats was carried out in two parts for different parameters. Among five different groups of rats in each part, a positive control group received α-lipoic acid in 0.5% CMC while test groups received 200 mg/kg and 400 mg/kg of aqueous extract of B. diffusa orally for 10 days. Assessment of parameters such as blood urea nitrogen (BUN), serum creatinine level, kidney malondialdehyde (MDA), and glutathione (GSH) levels, kidney injury on histopathology. However, their results demonstrated that B. diffusa did not show significant improvement in PAH clearance, which was reduced due to gentamicin damage (Sawardekar and Patel, 2015). The results of the study showed that, aqueous extract of B. diffusa showed comparable results with positive control. 6.1.3.2. Antilithiatic Activity The growth-inhibition behaviour of struvite crystals by using in vitro single diffusion gel growth technique was studied. With increasing concentrations of herbal extract of B. diffusa, the number, dimensions, total mass, total volume, growth rate and depth of growth of struvite crystals were decreased (Chauhan et al., 2012). In ethylene glycol (0.75%) induced lithiatic rats, significant reduction in the elevated levels of calcium, phosphate, uric acid and oxalate ions in urine was observed on oral administration of alcoholic extract of roots (250 mg/kg b.w. and 500 mg/kg b.w.) of B. diffusa. The extract elevated the levels of magnesium in urine which acts as an inhibitor of crystallization. In this study, cycstone as a standard drug (at high dose of 750mg/kg b.w.) was used for comparative efficacy (Chitra et al., 2012). 6.1.4. Antioxidant activity Since B. diffusa can contains high tannins and phenols as well as flavonoids, many studies on antioxidant potential of B. diffusa have been reported (Gopal et al., 2010; Olaleye et al., 2010; Vaghasiya et al., 2011). Antioxidant activity guided chromatographic fractionation led to the isolation of eupatilin-7-O-α-rhamnosyl(1→2)α-rhamnosyl(1→6)-β-D-galactopyranoside, a non-catechol group flavone and two catechol group flavonol glycosides viz., quercetin-3-O-α-L-rhamnosyl(1→6)-βgalactopyranoside and quercetin-3,7-di-O-glucoside (Fig. 2) (Joshi et al., 2010). Ethanolic extract of

32

B. diffusa scavenged 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals with reduction potential of 0.65 ± 0.02 mg/g ascorbic acid (Olaleye

et al., 2010). An electron spin resonance (ESR)-guided

fractionation of the methanolic extract of B. diffusa led to the isolation of boeravinone D, boeravinone G and boeravinone H which showed remarkable radical-scavenging activity (Aviello et al., 2011). When they compared chemical structure of these three compounds, boeravinone G showed higher antioxidant activity remarkably at nanomolar range due to the absence of methyl group at position 10 (Fig. 2). This study further demonstrated that MAP kinase and NF-kB pathways seem to be involved in the antioxidant effect of boeravinone G and it might be useful in reactive oxygen species (ROS)mediated injuries (Aviello et al., 2011). In African tranditional medicine, the redder stems of B. erecta are preferred over less red. Led by this, a study evaluated the antioxidant potential of redder stem bark of B. erecta and proved that betacyanins (which gives a characteristic red colour to stems) are more antioxidant than phenolic compounds of the stem (Hilou et al., 2013). In another study, different extracts of B. elegans have scavenged the DPPH and reduced the ferric ion. This study concluded that the phenolic content of the extracts and antioxidant activity has positive correlation (Sadeghi et al., 2014). Similarly, polyphenolic concentration in B. procumbens found to be responsible for its marked antioxidant potential when assayed through various antioxidant parameters (Bokhari and Khan, 2015a). Very recently, Boerhavia diffusa, at a dose of 100 µg/ml, protected oxidative damage against quinolinic acid (QA), 3-nitropropionic acid (NPA), sodium nitroprusside (SNP), and Fe (II)/EDTA complex induced oxidative stress in rat brain homogenates. The study also demonstrated that B. diffusa can protect hydroxyl radical induced DNA damage in the tissues (IC50 = 38 91 ± 0 12 μg/ml) (Ayyappan et al., 2015). 6.1.5. Hepatoprotective activity Brazilian folk medicines as well as Indian medicinal practitioners have been using B. diffusa and B. erecta for various hepatic disorders. So, it is much of interest for researchers to investigate their hepatoprotective potential. Pre-treatment of the ethanolic extract of B. diffusa (100, 200, 300 and 400 mg/kg) once daily for 7 consecutive days in acetaminophen-induced liver damage rats decreased activity of serum enzymes which was elevated by acetaminophen. Also, acetaminophen induced oxidative stress which was counteracted by B. diffusa extract. The negative control in this study received physiological saline but a positive control seems to be lacking (Olaleye et al., 2010). Very recent study suggested that the aqueous extracts of B. diffusa when orally administered at the doses of 250 mg/kg b.w. /day and 500 mg/kg b.w. /day for 4 days decreased the elevated levels of alanine transferases (ALT), serum asparate transferases (AST), alkaline phosphatase (ALP) and serum albumin against CCl4 induced liver toxicity in albino rats. The positive control in the study was silymarin (50 mg/kg/ b.w.) (Beedimani et al., 2015). 6.1.6. Antiviral property 33

From above hepatoprotective activities it might be suggested that B. diffusa has curative role in the liver damage. According to modern studies, B. diffusa showed a potential to cure infectitious hepatitis by antiviral mechanism. In a study, 90% EtOH root extract (5 mg/ml) of B. diffusa showed antiviral potency by inhibiting surface antigen as well as inhibition of HBV (hepatitis B virus) DNA polymerase (Kannan et al., 2011). Additionally, 200 µg/ml of B. diffusa induced a proinflammatory Th1 antiviral cytokine i.e. IFN-γ in peripheral blood mononuclear cells (PMBC). For this cytokinin induction activity, PHA (5 µg/ml) was used as positive control. All these results further concluded that B. diffusa contains anti-HBV substance(s) but exact mechanism of action need to be elucidated (Kannan et al., 2011). Recently, a supportive study to antiviral principles in Boerhavia genus came from moderate anti-HIV integrase activity of quercetin-3-O-rutinoside (IC50 = 10 µg/ml) and isorhamnetin-3-O-rutinoside (IC50 = 22 µg/ml) isolated from stem bark of B. erecta (Nugraha et al., 2015). Effectiveness of Boerhavia species have been already reported against plant viruses (Awasthi and Verma, 2006) but the above studies indicated their potential against human viruses which would be of certain interest for evaluation of molecular mechanism and production of antiviral compounds from Boerhavia. 6.1.7. Anticancer effect Indian Ayurveda has mentioned the use of B. diffusa in the treatment of tumours which may have driven the researchers to explain its anticancer potential through different mechanisms. Treatment of punarnavine, an active component from B. diffusa resulted in the presence of apoptotic bodies and DNA fragmentation in B16F-10 melanoma cells in a dose dependent manner. The study demonstrated that punarnavine induces apoptosis via activation of p53 induced caspase-3 mediated pro-apoptotic signalling and suppression of NF-κB induced Bcl-2 mediated survival signalling (Manu and Kuttan, 2009a). Also, extracts from B. diffusa root showed that a methanol: chloroform fraction at a concentration of 200 μgL−1 signiﬁcantly reduced cell proliferation with visible morphological changes in HeLa cells after 48 hrs of exposure. Cell cycle analysis suggested that antiproliferative effect of B. diffusa could be due to inhibition of DNA synthesis in HeLa cells (Chopra et al., 2011). In another study, higher doses of phytoproteins from B. diffusa have shown anticarcinogenic potential in MCF-7 cell line possessing the membrane receptor for estrogens and androgen (Singh et al., 2012). Latest findings suggest that, punarnavine at 50 µM inhibited in vitro MMP-2 and MMP-9 expression in HUVECs and neovascularisation in sponge implant assay. It also showed in vivo anticancer potential by decreasing ascitic fluid volume by 60.94% and tumour volume by 86.40% in Ehrlich ascites model (Saraswati et al 2013). The study also demonstrated its role in anti-angiogenesis by inhibiting VEGF expression as evident from RT-PCR, ELISA and Western blotting assay (Saraswati et al., 2013). 6.1.8. Cardiovascular effects 34

Ethanolic extract of B. diffusa (20 μg/mL) found to protect 5 µM arsenic trioxide (ATO) -induced cardiotoxicity in H9c2 Myoblasts. A decreased activity of lactate dehydrogenase (6 61 ± 1 97 μU/mL, respective control group: 16 15 ± 1 92 μU/mL), reduced oxidative stress, reduced calcium influx and organelle damage marked the protective effect (Vineetha et al., 2013). In one more study, ethanolic extract of B. diffusa was shown to have protective effect against mitochondrial dysfunction in angiotensin II induced hypertrophy in H9c2 cardiomyoblast cells. The study demonstrated that activities of aconitase and thioredoxin reductase lowered due to hypertrophy and were increased by ethanolic extract of B. diffusa. Further, the extract significantly prevented the generation of intracellular ROS and mitochondrial superoxide radicals. It has protected the mitochondria by preventing dissipation of mitochondrial transmembrane potential (ΔΨm), opening of mitochondrial permeability transition pore (mPTP) and mitochondrial swelling. The extract also enhanced the activities of respiratory chain complexes and oxygen consumption rate in embryonic rat heart-derived H9c2 cell line (Prathapan et al., 2014). 6.1.9. Anti-inflammatory activity As described earlier, traditional use of Boerhavia in asthma and inflammatory disorders led to investigate its potential in these ailments. 95% EtOH extract of B. diffusa roots at the doses of 100 mg/kg, 200 mg/kg and 400 mg/kg showed protection against preconvulsive dyspnoea in histamine aerosol exposed experimental animals suggesting anti-histaminic activity. They further concluded that the activity may be due to H1-receptor blocking or bronchodilation (Suralkar et al., 2012). Boeravinone B, a rotenoid isolated from roots of B. diffusa showed significant in vivo antiinflammatory activity (56.6% at 50 mg/ kg), better than the positive control ibuprofen (43.52% and 50.40% at 50 and 100 mg/kg po, respectively) in female Sprague−Dawley rats (Bairwa et al., 2013a). ‘Punarnavasava’ is an Ayurvedic formulation mainly contains B. diffusa which inhibited carrageenan-induced paw edema in rats (Gharate and Kasture, 2013). Nitric oxide (NO) is a potent pleiotropic inhibitor of physiological processes such as smooth muscle relaxation, neuronal signaling and platelet aggregation. Ethanolic extract showed nitric oxide inhibition (IC50 = 370.47 ± 7.33) and scavenged the nitric oxide free radicals in vitro which led to conclusion that B. diffusa may have role in free radical mediated chain reactions in inflammation (Muthu et al., 2014). The same extract inhibited protein denaturation (IC50 = 356.29 ± 1.46) and proteinase activity (IC50 = 348.84 ± 1.40) (Muthu et al., 2014). Furthermore, the study demonstrated that ethanolic extract reduced the thickness of paw volume in both carrageenan induced inflammation and cotton pellet induced granuloma, showing in vivo anti-inflammatory effect (Muthu et al., 2014). Very recent study has isolated five new rotenoids viz., boeravinones P, boeravinones Q, boeravinones R, boeravinones S, boeravinones M and four known viz., boeravinone A, 10-O-demethylboeravinone C, boeravinone B and boeravinone E as anti-inflammatory agents from 70% ethanolic extract of B. diffusa (Bairwa et al., 2013a; Bairwa et al.,

35

2013b). Among these, boeravinone S (40 μM) showed highest in vitro COX-1 (IC50 = 21.1 μM) and COX-2 (IC50 = 26.7 μM) inhibitory activities. 6.1.10. Anticonvulsant activity In Nigerian folk medicine, the B. diffusa has been known in treatment of epilepsy and hence study has been carried to elucidate its usefulness in convulsions. Anti-convulsant activity of methanolic extract of B. diffusa roots (1000, 1500 and 2000 mg kg−1, intraperitoneally (i p )) and liriodendrin-rich fraction (10, 20 and 40mg kg−1, i.p.), chloroform fraction (20mg kg−1, i p ) and phenolic compound fraction (1 mg kg−1, i p ) were studied in pentylenetetrazol (PTZ) (75 mg kg−1, i p )-induced seizures. The crude methanolic extract of B. diffusa and only its liriodendrin-rich fraction showed a dose-dependent protection against PTZ-induced convulsions. The observed anticonvulsant activity was due to calcium channel antagonistic action of the liriodendrin-rich fraction (Kaur and Goel, 2011). 6.1.11. Antibacterial activity Nowadays, researchers are focusing on plant extracts and their metabolites as antimicrobials because of prevalence of multi-drug resistant bacteria and their low susceptibility to antibiotics. Apu et al. (2012) have noticed that the methanolic extract of Boerhavia diffusa was possessed antimicrobial activity against three pathogenic organisms namely, Staphylococcus aureus (Gram positive), Shigella dysenteriae (Gram negative) at 1000 µg/disc of extract. The study used ciprofloxacin (5 µg/disc) as positive control. The ethanolic extract of whole plant of Boerhavia diffusa showed antimicrobial activity against bacterial strains Bacillus subtilis UC564, Staphylococcus aureus 15 ML296, Staphylococcus aureus ML329 and Salmonella typhi DI at 2000 µg/ml. Ofloxacin was used as positive control (Das, 2012). 7. Toxicity studies While the Boerhavia species offer cures for variety of ailments, its use in traditional medicine as emetic endorses toxicity. Boerhavia diffusa L., as of 2007, has been enlisted in US govt. FDA poisonous plant database (www.accessdata.fda.gov/scripts) which seems to be primarily sourced from different books (Bessey, 1902; Dastur, 1962; McGuffin et al., 2000; Merrill, 1943). According to one such book, roots of B. diffusa affects kidney if used as food (Merrill, 1943). However, scientific study suggest no any observable adverse effect on foetal development after daily administration of 250 mg/kg b.w. of ethanolic root extract in rats throughout pregnancy (Singh et al., 1991). A range from 500, 1000, 2000 mg/kg b.w. of aqueous extract of B. diffusa leaves given orally to albino mice and rats did not affect both the absolute and relative organ weights between the control and the test group. The liver enzymes and haematological parameters were statistically equal in all the groups and

36

suggested LD50 value more than 2000 mg/kg b.w. (Orisakwe et al., 2003). Nevertheless, decoction of stem bark of B. erecta was observed to have low acute toxicity with LD50 value of 2148 mg/kg b.w. in male mice (Hilou et al., 2006). This can be correlated with the traditional use of Boerhavia species as emetic when taken in large dose. Also, 80% ethanol extract of aerial parts of B. elegans have shown very low toxicity with LD50 value of 1020 µg/ml when screened with brine shrimp (Artemia salina) toxicity test (Ramazani et al., 2010). Conversely, Boerhavia chinensis has shown potential neurotoxicity at LD50 value of ˃1000 mg/kg b.w. in mice of either sex weighing 15-20gms (Dhawan et al., 1980). In short-term toxicity test, the aqueous leaf extract of Boerhavia erecta at the dose of 1000 mg/kg/day for 28 days affected glucose level in male and female Wistar rats. At the same time, the sub chronic toxicity of the same extract at three doses of 100 mg/kg/day, 300 mg/kg/day and 1000 mg/kg/day, orally administered for 90 days, did not result in any mortality (Lagarto et al. 2011). A study claimed that crude methanolic extract of B. repens found to be potentially toxic when tested at different concentrations (0.78125, 1.5625, 3.125, 6.25, 12.50, 25, 50, 100, 200 and 400 μg/ml) against Artemia salina (brine shrimp lethality assay). The LC50 value (based on Log C) for B. repens observed in this study was 4.19 µg/ml compared to vincristine sulphate as the positive control (LC50 = 0.840 μg/ml based on Log C) (Rahman et al., 2014). 8. Conclusion From above studies, it is concluded that the genus Boerhavia has long been used as traditional medicine worldwide most notably in liver disorders, inflammation, urinary disorders, gastro-intestinal problems, malaria, asthma and microbial infections. Many studies reported flavonoids as major bioactive compounds from this genus and their number seems to be increased with advent of research. Various rotenoids are isolated and primarily well investigated for anticancer, anti-inflammatory and antioxidant potential. Several studies are repeatedly published on antidiabetic, hepatoprotective and nephroprotective effects of Boerhavia species. Latest findings have shown interest in obtaining antiviral and anti-inflammatory principles from genus Boerhavia. However, there are some problems which need to be addressed (i) Although most studies have focused on evaluating the bioactive potential of B. diffusa but other species of Boerhavia still need attention. For this to achieve, research also need to be directed in setting up proper tools for effective discrimination of Boerhavia species. (ii) Many studies claimed the bioactive potential of phytochemicals from genus Boerhavia. However, further research is needed on the mechanism of action or pharmacodynamics of phytochemical compounds to clarify their bioefficacy as well as feasibility for commercial drug formulation. (iii) Because of inadequately characterized data, various pharmaceutical activities reported for quinolizidine alkaloid i.e. punarnavine are erratic. iv) Clinical studies should have priority for compounds with well established pharmaceutical activities. (v) Leaves and roots are mainly exploited for phytochemical and pharmaceutical investigation. Study of other parts such as seeds, flowers and stems are necessary for effective utilization of these species. (vi) Though few reports disprove the 37

toxicity of B. diffusa through various assays, toxicity studies must be carried out in all the members of Boerhavia as there are few reports of potential toxicity in B. repens and B. chinensis. Acknowledgement One of the authors, Mr. Kapil Suresh Patil is thankful to UGC, New Delhi for financial support to this work under the Research Fellowship in Science for Meritorious Students (RFSMS) (sanctioned vide letter No F. 4-1/2006 (BSR)/7-137/2007 (BSR) dated June 26, 2012). Authors gratefully acknowledge financial assistance towards research facility at School of Life Sciences, NMU, Jalgaon from UGC, New Delhi under SAP-DRS programme and DST, New Delhi under FIST programme. References Abbasi, A.M., et al., 2009. Medicinal plants used for the treatment of jaundice and hepatitis based on socio-economic documentation. African Journal of Biotechnology 8(8), 1643-1650. Agarwal, R.R., Dutt, S.S., 1936. Chemical examination of Punarnava or Boerhavia diffusa Linn.II. Isolation of an alkaloid Punarnavine. Chemical Abstract 30, 44-54. Ahmed, B., Yu, C.P., 1992. Borhavine- A dihydroisofuranoxanthone from Boerhaavia diffusa. Phytochemistry 31, 4382-4384. Andesina, S.K., 1982. Studies on some plants used as anticonvulsants in Amerindian and African traditional medicine. Fitoterapia 53, 147-162. Anjalam, A., Elangomathavan, R., Premalatha S., 2014. Exploration of medicinal plants used by the Malyali tribes of Kolli hills, Tamilnadu, India. International Journal of Research in Ayurveda and Pharmacy 5(2), 201-204. Apu, A.S., Liza, S.M., Jamaluddin, A.T.M., Md. A.H., Saha, R.K., Rizwan, F., Nasrin, N., 2012. Phytochemical screening and in vitro bioactivities of the extracts of aerial part of Boerhavia diffusa Linn. Asian Pacific Journal of Tropical Biomedicine 2(9), 673-678. Austin, D.F., 2010. Baboquivari Mountain Plants: Identification, Ecology, and Ethnobotany. The University of Arzona Press, USA, pp. 177. Aviello, G., Jasna, M., Canadanovic-Brunet, Milic, N., Capasso, R., Fattorusso, E., TaglialatelaScafati O., Fasolino L., Izzo A.A., Borelli, F., 2011. Potent antioxidant and genoprotective effects of Boeravinone G, a rotenoid isolated from Boerhaavia diffusa. PLoS ONE 6(5), e19628. doi: 10.1371/journal.pone.0019628. Awasthi, L.P., Verma, H.N., 2006. Boerhaavia diffusa- a wild herb with potent biological and antimicrobial properties. Asian Agri-History 10(1), 55-68. Ayyappan, P., Palayyan, S.R., Gopalan, R.K., 2015. Attenuation of oxidative damage by Boerhaavia diffusa L. against different neurotoxic agents in rat brain homogenate. Journal of Dietary Supplements. doi: 10.3109/19390211.2015.1036186. Bairwa, K., Singh, I.N., Roy, S.K., Grover, J., Srivastava, A., Jachak, S.M., 2013a. Rotenoids from Boerhaavia diffusa as potential anti-inflammatory agents. Journal of Natural Products 76, 1393– 1398. Bairwa, K., Singh, I.N., Roy, S.K., Grover, J., Srivastava, A., Jachak, S.M., 2013b. Correction to rotenoids from Boerhaavia diffusa as potential anti-inflammatory agents. Journal of Natural Products 76, 2364. Bairwa, K., Srivastava, A., Jachak, S.M., 2014. Quantitative analysis of boeravinones in the roots of Boerhaavia diffusa by UPLC/PDA. Phytochem. Anal. 25 (5), 415-420. doi:10.1002/pca.2509. Beedimani, R.S., Jeevangi, S.K., 2015. Evaluation of hepatoprotective activity of Boerhaavia diffusa against carbon tetrachloride induced liver toxicity in albino rats. International Journal of Basic & Clinical Pharmacology 4(1), 153-158. doi: 10.5455/2319-2003.ijbcp20150230. 38

Beegum, G.R.J., Beevy, S.S., Sugunan, V.S., 2014. Nutritive and anti-nutritive properties of Boerhavia diffusa L. J. Pharma. Phytochem. 2 (6), 147-151. Belkacem, A.A., Macalou, S., et al., 2007. Nonprenylated rotenoids, a new class of potent breast cancer resistance protein inhibitors. Journal of Medicinal Chemistry 50, 1933-1938. Bep Oliver-Bever., 1986. Medicinal plants in tropical West Africa. Cambridge University Press, p. 196. Bessey, C.E., 1902. Preliminary account of the plants of Nebraska which are reported to be poisonous or suspected of being so. Nebraska Agriculture Bd. Annual Reports 1901, 95-129. Bharali, R., Azad, M.R.H., Tabassum J., 2003. Chemopreventive action of Boerhaavia diffusa on DMBA–induced skin carcinogenesis in mice Indian Journal Physiology and Pharmacology 47 (4), 459–464. Bhatia, V., Kinja, K., Bishnoi, H., Savita, S., Gnaneshwari, D., 2011. Antidiabetic activity of the alcoholic extract of the arial part of Boerhaavia diffusa in rats. Recent Research in Science and Technology 3(7) , 04-07. Bhope, S.G., Gaikwad, P.S., Kuber, V.V., Patil, M.J., 2013. RP-HPLC method for the simultaneous quantitation of boeravinone E and boeravinone B in Boerhaavia diffusa extract and its formulation. Natural Product Research 27(6), 588–591. Bhope, S.G., Ghosh, V.K., Kuber, V.V., Patil, M.J., 2011. Rapid microwave-assisted extraction and HPTLC- photodensitometric method for the quality assessment of Boerhaavia diffusa L. Journal of AOAC International 94(3), 795-802. Bokhari, J., Khan, R.M., 2015a. Evaluation of anti-asthmatic and antioxidant potential of Boerhavia procumbens in toluene diisocyanate (TDI) treated rats. Journal of Ethnopharmacology 172, 377– 385. Bokhari, J., Khan, R.M., Haq, I.U., 2015b. Assessment of phytochemicals, antioxidant, and antiinflammatory potential of Boerhavia procumbens Banks ex Roxb. Toxicology and Industrial Health 1–11. doi: 10.1177/0748233714567183. Bossard, E., 1996. La médecine traditionnelle au centre et à l'ouest de l'Angola. Ministério da ciênciae da tecnologia. Instituto de investigaçâo cientifica tropical. Lisboa p. 531. Bouquet, A., Debray M., 1974. Plantes médicinales de la Côte d'Ivoire. Travaux et Documents de l'O.R.S.T.O.M., Paris, pp. 232. Braga, R., 1960. Plantas do Nordeste, Especialmen~e do Ceark, Imprensa otlicial, Fortaleza, Cear and Brazil, pp. 406. Buchanan, B.B., Gruissem, W., Jones, R.L., 2015. Biochemistry and molecular biology of plants. John Wiley & Sons, UK. Second edition, pp.1133. Chauhan, C.K., Joshi M.J., Vaidya, A.D.B., 2012. Antilithiatic activity of Boerhaavia diffusa Linn, Commiphora wightii and Rotula aquatica lour on urinary type struvite crystals. Abstract proceedings P-033 ISP. Chitra, V., Gowrk, I., Udayasri, N., Antilithiatic activity of alcoholic extract of Boerhaavia diffusa roots on ethylene glycol induced lithiasis in rats. International Journal of Pharmacy and Pharmaceutical Sciences 4(2), 149-153. Chopra, GL., Angiosperms: Systematics and Life Cycle. S. Nagin & Co., Jalandhar, Punjab, India; 1969; p. 361-365. Chopra, M., Srivastava, R., Saluja, D., Dwarakanath, B.S., et al., 2011. Inhibition of human cervical cancer cell growth by ethanolic extract of Boerhaavia diffusa Linn. (Punarnava) root. Evidence Based Complementary and Alternative Medicine 13. Chopra, R.N., Nayar, S.L., Chopra, I.C., 1956. Glossary of Indian Medicinal Plants. Council of Scientific and Industrial Research (CSIR), New Delhi, India, p. 39. Chou, F.S., Liu, H.Y., Sheue, C.R., 2004. Boerhavia erecta L. (Nyctaginaceae), a new adventive plant in Taiwan. Taiwania 49(1), 39-43. Chude, M.A., Orisakwe, O.E., Affone, O.J., Gamaniel, K.S., Vongtau, O.H., Obi, E., 2001. Hypoglycemic effect of the aqueous extract of Boerhaavia diffusa leaves. Indian Journal of Pharmacology 33, 215–216. Crombie L., 1984. Rotenoids and their Biosynthesis. Natural Product Reports 1(1), 3-19. doi:10.1039/NP9840100003. 39

Crombie, L., Josephs, J.L., Cayley, J., Larkin, J., Weston, J.B., 1992. The rotenoid core structure: Modifications to define the requirements of the toxophore. Bioorganic Medicinal Chemistry Letters, 2, 13-16. Cruz, G.L., 1995. Dicionario Das Plantas Uteis Do Brasil. 5th Edition. Bertrand, Rio de Janeiro, Brazil. Daniel M. Medicinal Plants: Chemistry and Properties. Science Publishers, Enfield, NH, USA, 2006. pp. 44. Dastur, J. F., 1962. Medicinal Plants of India and Pakistan. DB Taraporevala Sons. Bombay, pp. 212. Dev, S., 2006. A selection of prime Ayurvedic plant drugs-An ancient modern concordance. Anamaya publishers, New Delhi, pp. 501. Dhawan, B.N., Dubey, M.P., Mehrotra, B.N., Tandon, J.S., 1980. Screening of Indian plants for biological activity: part IX. Indian Journal of Experimental Biology 18, 594-602. Dhingra, D., Valecha, R., 2014. Punarnavine, an alkaloid isolated from ethanolic extract of Boerhaavia diffusa Linn. reverses depression-like behaviour in mice subjected to chronic unpredictable mild stress. Indian Journal of Experimental Biology 52(8), 799-807. Do, T.M.L., et al., 2013a. New rotenoids and coumaronochromonoids from the aerial part of Boerhaavia erecta. Chem. Pharm. Bull. 61(6): 624–630. Do, T.M.L., et al., 2013b. New derivatives from the aerial parts of Boerhaavia diffusa L. (Nyctaginaceae). Phytochemistry Letters 6 (4), 544-551. Donald, H.P., 2012. CRC world dictionary of medicinal and poisonous plants. Umberto Quattrocchi, F.L.S. CRC Press, Taylor & Francis Group, p-613. Douglas, N.A., Manos, P.S., 2007. Molecular phylogeny of Nyctaginaceae: Taxonomy, biogeography and characters associated with a radiation of xerophytic genera in North America. American Journal of Botany 96, 856-872. Edeoga, H.O., Ikem, I.C., 2002. Structural morphology of the pollen grains in three Nigerian Species of Boerhavia L. New Botany 29, 89-95. Fadeyi, M.O., Adeoye, A.O., Olowokudejo J.D., 1989. Epidermal and phytochemical studies in the genus Boerhavia (Nyctaginaceae) in Nigeria. International J. Crude Drug Res. 27 (3), 178-184. Fathima, K.A., 2012. Nephroprotective effect of Eupalitin-3-O-β-D-galactopyranoside in Koi carp (Cyprinus carpio) Journal of Pharmacy Research 5(5): 2590-2592. Ferreres, F., Sousa, C., et al., 2005. Characterisation of the phenolic profile of Boerhaavia diffusa L by HPLC-PAD-MS/MS as a tool for quality control. Phytochemical Analysis 16, 451–458. Fosberg, F., 1978. Studies in the genus Boerhavia L. (Nyctaginaceae), 1-5. Smithsonian Contributions to Botany, number 39, Smithsontan Institution Press, Washington, p. 1-20. Gharate, M., Kasture, V., 2013. Evaluation of anti-inflammatory, analgesic, antipyretic and antiulcer activity of Punarnavasava: an Ayurvedic formulation of Boerhavia diffusa. Oriental Pharmacy of Experimental Medicine 13, 121–126. Goel, R.K., Singh, D., Lagunin, A., Poroikov, V., 2011. PASS-assisted exploration of new therapeutic potential of natural products. Medicinal Chemistry Research 20, 1509–1514. doi:10.1007/s00044-010-9398-y. Gopal, T.K., Harish, G., Chamundeeswari, D., Reddy, C.U., 2010. In-vitro anti-oxidant activity of roots of Boerhaavia diffusa Linn. Research Journal of Pharmaceutical Biology and Chemical Sciences 1(4), 782-788. Guessan N’ K , Kadja, B , ZirihiI, G N , Traoré, D , Aké-Assi, L., 2009. Screening phytochimique de quelques plantes médicinales ivoiriennes utilisées en pays Krobou (Agboville, Côte-d’Ivoire). Sciences & Nature 6 (1), 1 - 15. Gupta, D.R., Ahmed, B., 1984. A new C-methyl flavone from Boerhaavia diffusa Linn. roots. Indian Journal of Chemistry 23B, 682-684. Gupta, J., Ali, M., 1998. Chemical constituents of Boerhaavia diffusa Linn. roots. Indian Journal of Chemistry 37: 912-917. Hilou A., Nacoulma O.G., Guiguemde O.R., 2006. In vivo antimalarial activities of extracts from Amaranthus spinosus L. and Boerhaavia erecta L. in mice. Journal of Ethnopharmacology 103, 236–240.

40

Hilou, A., Millogo-Rasolodimby, J., Nacoulma, O.G., 2013. Betacyanins are the most relevant antioxidant molecules of Amaranthus spinosus and Boerhavia erecta. Journal of Medicinal Plants Research 7(11), 645-652. Hiruma, L.C.A., Gracioso, J.S., Bighetti, E.J., Germonsén, Robineou L., Souza, Brito, A.R., 2000. The juice of fresh leaves of Boerhaavia diffusa L. (Nyctaginaceae) markedly reduces pain in mice. Journal of Ethnopharmacology. 71, 267-274. Indhumathi, T., Shilpa, K., Mohandass, S., 2011. Evaluation of Nephroprotective role of Boerhaavia diffusa leaves against mercuric chloride induced toxicity in experimental rats. Journal of Pharmacy Research 4(6), 1848-1850. Jain, G.K., Khanna, N.M., 1989. Punarnavoside: A new antifibrinolytic agent from Boerhaavia diffusa Linn. Indian Journal of Chemistry 28(B), 163-166. Jayavelu, A., Natarajan, A., Sundaresan, S., Devi, K., Kumar S.B., 2013. Hepatoprotective Activity of Boerhavia diffusa Linn. (Nyctaginaceae) against Ibuprofen Induced Hepatotoxicity in Wistar Albino Rats. International Journal of Pharma Research and Review 2(4), 1-8. Jiofack, T., Ayissi, L., Fokunang, C., Guedje, N., Kemeuze, V., 2009. Ethnobotany and phytomedicine of the upper Nyong valley forest in Cameroon. African Journal of Pharmacy and Pharmacology 3(4), p 144-150. Joshi, N., Verma, D.L., 2012. Eupatilin-7-O-2,6-dirhamnosylgalactoside from the leaves of Boerhaavia diffusa Linn. Journal of Applied Chemistry 2(2), 13-17. Kadota, S., Lami, N., Tezuka, Y., Kikuchi, T., 1988a. Boeravinone A and B, new rotenoid analogues from Boerhaavia diffusa Linn. Chemical Pharmaceutical Bulletin 36, 834-836. Kadota, S., Lami, N., Tezuka, Y., Kikuchi, T., 1988b. Structure and NMR spectra of boeravenone C, a new rotenoid analogue from Boerhaavia diffusa Linn. Chem. Pharm. Bull. 36, 2289-2292. Kannan, M., Paramasivam, R., Chandran, P., Veerasami, V., Gnanasekaran, A., Shanmugam, A., Murugesan, K., 2011. Inhibition of hepatitis B virus DNA polymerase and modulation of Th1 & Th2 cytokine secretion by three Indian medicinal plants and its correlation with antiviral properties. Journal of Pharmacy Research 4(4), 1044-1046. Katewa, S.S., Chaudhari, B.L., Jain, A., 2004. Folk hebal medicines from tribal area of Rajasthan, India. Journal of Ethnopharmacology 92, 41-46. Kaur, M., Goel, R.K., 2011. Anti-convulsant activity of Boerhaavia diffusa: plausible role of calcium channel antagonism. Evidence Based Complimentary and Alternative Medicine 2011; 1-7. doi:10.1093/ecam/nep192. Khan, A., Thapliyal, R.P., Chauhan, S.K., 2012. Regulatory effect of Boerhaavia diffusa and black caraway oil on hepatic, lung and kidney antioxidants enzymes content in dmba-induced hypercholesterolemia in rats International Journal of Pharm. Pharm. Sciences 4, 384-392. Khare, C.P., 2004. Indian herbal remedies: rational western therapy, Ayurvedic and other traditional usage, botany. Springer Science & Business Media, pp. 104-105. Kokate, C.K., Purohit A.P., Gokhale, S.B., 2005. Pharmacognosy. 38th Edn., Nirali Prakashan, Pune, India, pp. 537-538. Krings, U., Berger, R.G., 2000. Antioxidant activity of some roasted foods. Food Chemistry 72, 223229. Krishna, V., Shanthamma, C., 2004a. Hepatoprotective activity of root extracts of Boerhaavia erecta L and B. rependa L. Indian Journal of Pharmaceutical Sciences 66(5), 667-670. Krishna, V., Shanthamma, C., 2004b. Boerhavia rependa-In vitro propagation and screening of hepatoprotective activity of leaves from field grown plants and leaf calli grown under in vitro. Journal of Tropical Medicinal Plants 5(2), 237-243. Lagarto, A., et al. 2011. Acute, short-term, and subchronic oral toxicity of Boerhaavia erecta L extract and Cucurbita pepo L seed oil. International J. Pharma. Toxi. Sci. 3, 9-21. Lami, N., Kadota, S., Kikuchi, T. 1992. Constituents of the roots of Boerhaavia diffusa Linn. IV. Isolation and structure determination of boeravinones D, E and F. Chemical Pharmaceutical Bulletin 39, 1863-1865. Levin, R.A., Raguso, R.A. & Mcdade, L.A. 2001. Fragrance chemistry and pollinator affinities in Nyctaginaceae. Phytochemistry 58: 429–440. Li, J , Li, H , Kadota, S , Νamba, T , 1996 Effects on cultured neonatal mouse Calvaria of the flavonoids isolated from Boerhaavia repens. Journal of Natural Products 59, 1015-1018. 41

Ma, W, et al., 2014. A review on astringency and bitterness perception of tannins in wine, Trends Food Sciences Technology 40 (1), 6-19. doi:10.1016/j.tifs.2014.08.001. Mahesh, A.R., Ranganath, M.K., Harish Kumar D.R., 2011. Enrichment of flavonoids from the methanolic extract of Boerhaavia diffusa roots by partitioning technique. Research Journal of Chemical Sciences 3(1), 43-47. Manu, A.K., Kuttan, G., 2009a. Punarnavine induces apoptosis in B16F-10 melanoma cells by inhibiting NF-kB signalling. Asian Pacific Journal of Cancer Prevention 10, 1031-1038. Manu, K.A., Kuttan, G., 2009b. Immunomodulatory activities of Punarnavine, an alkaloid from Boerhaavia diffusa. Immunopharmacology and Immunotoxicology 31(3):377-387. Maurya, R., Sathiamoorthy, B., Mundkinajeddu, D., 2007. Flavonoids and phenol glycosides from Boerhavia diffusa. Natural Product Research 21(2): 126–134. McGuffin, M., Kartesz, J.J. Leung, A.Y., Tucker, A.O., 2000. Herbs of commerce, Second Edition, American Products Association, pp. 421. Merrill E.D., 1943. Emergency food plants and poisonous plants of the islands of the Pacific. Washington, D.C., U.S. Govt. PrInternational Office, 1943, pp. 55. Misra A.N., Tiwari, H.P. 1971. Constituents of roots of Boerhaavia diffusa. Phytochem. 10, 33183319. Mitra, R., Gupta, R.C., 1997. Punarnava – An Ayurvedic drug of repute In: Applied Botany Abstracts, Economic Botany Information Service, National Botanical Research Institute, Lucknow, Uttar Pradesh, India 17(3), p. 209-227. Mundkinajeddu, D., Manahas, L.R., Maurya, R., Handa, S. S., 2003. Process of isolation of eupalitin from Boerhaavia diffusa. United State Patent No. US 20030175373 A1, Publication date: Sep 18, 2003. Mungantiwar, A.A., Nair, A.M., Shinde, U.A., Dikshit, V.J., et al., 1999. Studies on the immunomodulatory effects of Boerhaavia diffusa alkaloidal fraction. Journal of Ethnopharmcology 65(2), 125–131. Muthu, S., Sivaganesh, M., Ashir, S., Tamilselvan, A., Sakeer, S., 2014. Anti-inflammatory effect of ethanolic extract of Boerhavia diffusa leaves in Wistar rats. Malaysian Journal of Bioscience 1(2), 76–85. Muzila, M., 2008. Boerhavia diffusa L, in: Schmelzer G.H., Gurib-Fakim A., (Editors). Plant Resources of Tropical Africa 11(1), Medicinal plants, PROTA Foundation, Wageningen, Netherlands / Backhuys Publishers, Leiden, pp. 117-121. Nalamolu, R.K., Boini, K.M., Nammi, S., 2004. Effect of chronic administration of Boerhaavia diffusa Linn leaf extract on experimental diabetes in rats. Tropical Journal of Pharmaceutical Research 3 (1), 305-309. Nugraha, A., 2010. Bioactive compounds from Boerhavia erecta L.: an African medicinal plant, Master of Science-Research Thesis, School of Chemistry, University of Wollongong, http://ro.uow.edu.au/theses/3674. Nugraha, A.S., Hilou, A., Vandegraaff, N., Rhodes, D.I., Haritakun R., Keller P.A., 2015. Bioactive glycosides from the African medicinal plant Boerhavia erecta L., Natural Product Research DOI: 10.1080/14786419.2015.1013470. Ojewole, A.J.O., Andesina, S.K., 1985. Isolation, identification and some cardiovascular actions of a purine nucleoside from the roots of Boerhaavia diffusa. Fitoterapia 56(1), 31-36. Olaleye M.T., Akinmoladun A.C., Ogunboye, A.A., Akindahunsi, A.A., 2010. Antioxidant activity and hepatoprotective property of leaf extracts of Boerhaavia diffusa Linn. against acetaminophen-induced liver damage in rats. Food and Chemical Toxicology 48(8-9), 22002205. Orisakwe, E.O., Afonne, J.O., Chude, A.M., Obi, E., Dioka, E.C., 2003. Sub-chronic toxicity studies of the aqueous extract of Boerhaavia diffusa leaves. Jouranl of Health Science 2003; 49(6), 444447. Pathak D., Alam K., Rohilla H., Rai A.K., Agrawal A., 2012. Phytochemical investigation of Boerhavia diffusa and Andrographis paniculata: a comparative study. International Journal of Pharm Pharmc Sci 4(4), 250-251.

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Patil, K.S., Bhalsing, S.R., 2015. Efficient micropropagation and assessment of genetic fidelity of Boerhaavia diffusa L- High trade medicinal plant. Physiology and Molecular Biology of Plants 21(3), 425–432. doi: 10.1007/s12298-015-0301-7. Pereira, D.M., Faria, J., Gaspar, L., Valentao, P., Andrade, P.B., 2009. Boerhaavia diffusa: metabolite profiling of a medicinal plant from nyctaginaceae. Food and Chemical Toxicology 47, 21422149. Petrus, A.J.A., Siva S., Hemalatha, Suguna, G., 2012. Isolation and characterisation of the antioxidant phenolic metabolites of Boerhaavia erecta L. leaves. Journal of Pharmaceutical Sciences and Research 4(7), 1856 – 186. Prathapan, A., Prabhakaran, V., Vineetha, Raghu, K.G., 2014. Protective effect of Boerhaavia diffusa L. against mitochondrial dysfunction in Angiotensin II induced hypertrophy in H9c2 cardiomyoblast cells. PLoS ONE 9(4), e96220. doi: 10.1371/journal.pone.0096220. Pullaiah, T., 2006. Encyclopaedia of World Medicinal Plants, Volume 1. Regency publications, New Delhi, India. p-337. Rahman, S.M., Alam, Md. M., Amin, M. R., Fakruddin, Md., Shahid-Ud-Daula, A.F.M., Siddiqui, R., 2014. Antimicrobial activity and brine shrimp toxicity of methanolic whole plant extract of Boerhavia repens L. (Family: Nyctaginaceae). International Journal of Phytopharmacy 4(6), 135-139. Rajeshwari, P., Krishnakumari, S., 2010. Boerhaavia erecta-A potential source for phytochemicals and antioxidants. Journal of Pharmaceutical Sciences and Research 2 (11), 728-733. Ramazani, A., Zakeri, S., Sardari, S., Khodakarim, N., Dinparas, Djadidt, N., 2010. In vitro and in vivo anti-malarial activity of Boerhavia elegans and Solanum surattense. Malaria Journal, 9, 124. Rameshkumar, S., Ramakritinan, C.M., 2013. Floristic survey of traditional herbal medicinal plants for treatments of various diseases from coastal diversity in Pudhukkottai District, Tamilnadu, India. Journal of Coastal Life Medicine 1(3), 225-232. doi:10.12980/JCLM.1.2013C664. Rivera, D., Allkin, R., Obón, C., Alcaraz F., Verpoorte R., Heinrich M., 2014. What is in a name? The need for accurate scientific nomenclature for plants. Journal of Ethnopharmcology 152, 393– 402. Robineau L.G., Soejarto, D.D., 1996. Tramil: a research project on the medicinal plant resouces of the Caribbean, in: Balick, M.J., Elisabetsky, E., Laird, S.A., (Eds.), Medicinal Resources of the Tropical Forest, Columbia University Press, New York, pp. 318–325. Sadeghi, Z., Kuhestani, K., Abdollahi, V., Mahmood, A., 2014. Ethnopharmacological studies of indigenous medicinal plants of Saravan region, Baluchistan, Iran. Journal of Ethnopharmacology 153, 111–118. Saraswati, S., Alhaider, A.A., Agrawal, S.S., 2013. Punarnavine, an alkaloid from Boerhaavia diffusa exhibits anti-angiogenic activity via downregulation of VEGF in vitro and in vivo. ChemicoBiological Interactions 206, 204–213. Sawardekar, S. B., Patel, T. C., 2015. Evaluation of the effect of Boerhavia diffusa on gentamicin induced nephrotoxicity in rats. Journal of Ayurveda and Integrative Medicine. 6(2), 95-103. Selvaraj D, Shanmughanandhan D, Sarma RK, Joseph JC, Srinivasan RV, Ramalingam S. 2012. DNA barcode ITS effectively distinguishes the medicinal plant Boerhavia diffusa from its adulterants. Genomics, Proteomics and Bioinformatics 10, 364–367. doi:10.1016/j.gpb.2012.03.002. Shah, G.M. and Khan, M.A., 2006. Common medicinal folk recipes of siran valley, Mansehra, Pakistan. Ethnobotanical leaflets 1, p.5. Shrivastava, N., Padhya, M.A., 1995. Punarnavine profile in the regenerated roots of Boerhaavia diffusa L from leaf segments. Current Science 68(6), 653–656. Singh, A., Singh, R.G., Singh, R.H., Mishra, N., Singh, N., 1991. An experimental evaluation of possible teratogenic potential in Boerhaavia diffusa in albino rats. Planta medica, 57(4), 315316. Singh, A.K., Najam, A., Pant, A.B., Awasthi, L.P., Verma, H.N., et al., 2012. Modulatory effect of phytoproteins isolated from roots of Boerhaavia diffusa and leaves of Clerodendrum aculeatum on neoplastic growth of human breast cancer cell line. Open Access Scientific Reports 1(9), 1-6. doi:10.4172/scientificreports.429.

43

Souza, J.M., Berkov, S., Santos, A.S., 2014. Improvement of friable callus production of Boerhaavia paniculata Rich and the investigation of its lipid profile by GC/MS. Anais da Academia Brasileira de Ciências, 86(3), 1015-1027. Spellenberg R.W., 2004. Boerhavia diffusa: in: Flora of North America, Editorial Committee eds. 2004. Oxford University Press, pp. 18. Stemmerik, J.F., 1964. Flora of Malesiana. Vol. 6. Groningen, The Netherlands: Wolters-Noordhoff Publishing. Stintzing, F.C., Kammerer, D., Schieber, A., Hilou, A., Nacoulma, O., Carle, R., 2004. Betacyanins and phenolic Compounds from Amaranthus spinosus L. and Boerhaavia erecta L. Zeitschrifts fur Naturforsch 59c: 1-8. Suralkar, A.A.,Verma, A.K., Kamble, R.D., Tayade, G.V., 2012. Pharmacological Evaluation of Anti-Histaminic activity of Boerhaavia diffusa. International Journal of Advanced Pharmaceutical Biology and Chemistry 1(4), 503-507. Suri, O.P., Kant, R., Jamwal, R.S., Suri, K.A., Atal, C.K., 1982. Boerhaavia diffusa, a new source of phytoecdysones. Planta Medica 44, 180-181. Tacchini, M., Spagnoletti, A., et al., 2015. Phytochemical profile and bioactivity of traditional ayurvedic decoctions and hydro-alcoholic macerations of Boerhaavia diffusa L. and Curculigo orchioides Gaertn. Natural Product Research 2015; doi:10.1080/14786419.2014.1003299. Tewari, D.N., Report of the task force on conservation & sustainable use of medicinal plants, Government of India Planning Commission, New Delhi. March – 2000. Thai, H.V., Kim, E., Kim, S.C., et al., Boerhavia diffusa L. ethanol extract suppresses inflammatory responses via inhibition of Src/Syk/TRAF6. Journal of Functional Foods 17, 476–490. Umamenaka, R., Vijayakumari, K., Senthilkumar, S., 2012. GC-MS analysis of Boerhaavia diffusa Linn leaf extract traditional valuable plant International Journal of Deccan Pharma and Life Sciences 3(2), 21-26. Vaghasiya, Y., Dave, R., Chanda, S., 2011. Phytochemical analysis of some medicinal plants from Western regions of India. Research Journal of Medicinal Plants 5(5), 567-576. Ved, D.K., Goraya, G.S., 2007. Demand and supply of medicinal plants in India. NMPB, New Delhi & FRLHT, Bangalore, India. pp. 15. Vineetha, V.P., Prathapan, A., Soumya, R.S., Raghu, K.G., 2013. Arsenic trioxide toxicity in H9c2 myoblasts-damage to cell organelles and possible amelioration with Boerhavia diffusa. Cardiovascular Toxicology 13(2), 123-37. doi: 10.1007/s12012-012-9191-x. Wabale, A.S., Petkar, A.S., 2005. Ethnomedicinal plants used against jaundice by the tribals of Akole taluka. Journal of Phytological research 18(2), 259-261. Wagner, H., Hikino, H., Farnsworth, N.R., 2012. Economic and Medicinal Plant Research. Academic press 3, pp. 109. Zagari, A., 1992. Medicinal Plants. 5th Edition. Tehran University Publications, Tehran, Iran Book, 4, pp. 969.

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