Persimmon (Diospyros kaki L.) leaves: a review on traditional uses, phytochemistry and pharmacological properties.

Journal of Ethnopharmacology ∎ (∎∎∎∎) ∎∎∎–∎∎∎

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Q1 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66

Contents lists available at ScienceDirect

Journal of Ethnopharmacology journal homepage: www.elsevier.com/locate/jep

Review

Persimmon (Diospyros kaki L.) leaves: A review on traditional uses, phytochemistry and pharmacological properties Chunyan Xie, Zhisheng Xie, Xinjun Xu n, Depo Yang School of Pharmaceutical Sciences, Sun Yat-Sen University, 510006 Guangzhou, PR China

art ic l e i nf o

a b s t r a c t

Article history: Received 9 August 2014 Received in revised form 7 January 2015 Accepted 7 January 2015

Ethnopharmacological relevance: Persimmon (Diospyros kaki L.) leaves, known as Shi Ye (in Chinese), have a long history as a Chinese traditional medicine for the treatment of ischemia stroke, angina, internal hemorrhage, hypertension, atherosclerosis and some infectious diseases, etc. Additionally, persimmon leaves could be used as healthy products, cosmetics and so on, which have become increasingly popular in Asia, such as Japan, Korea and China etc. Aim of the review: The present paper reviewed the ethnopharmacology, phytochemistry, analytical methods, biological activities and toxicology of persimmon leaves in order to assess the ethnopharmacological use and to explore therapeutic potentials and future opportunities for research. Materials and methods: Information on persimmon leaves were gathered via the Internet (using Google Scholar, Baidu Scholar, Elsevier, ACS, Pudmed, Web of Science, CNKI and EMBASE) and libraries. Additionally, information was also obtained from some local books. Results: Persimmon leaves have played an important role in Chinese system of medicines. The main compositions of persimmon leaves were flavonoids, terpenoids, etc. Scientific studies on extracts and formulations revealed a wide range of pharmacological activities, such as, antioxidative, hypolipidemic, antidiabetic, antibacterial, hemostasis activities and effects on cardiovascular system. Based on the pharmacological activities, persimmon leaves were widely used in clinic including treatment of cardiovascular disease, hemostasis, antibacterial, anti-inflammatory and beauty treatment. Conclusions: Persimmon leaves probably have therapeutic potential in the prevention and treatment for cerebral arteriosclerosis, diabetes, hypertension. It showed significant neuroprotection against ischemia/ reperfusion injury in vivo and in vitro. Moreover, it can regulate immune function and inhibite inflammation. Further investigations are needed to explore individual bioactive compounds responsible for these pharmacological effects in vitro and in vivo and the mode of actions. Further safety assessments and clinical trials should be performed before it can be integrated into medicinal practices. & 2015 Published by Elsevier Ireland Ltd.

Keywords: Diospyros kaki L. Persimmon leaves Flavonoids Terpenoids Cardiovascular effects Chemical compounds studied in this article: Quercetin (PubChem CID: 5280343) Kaempferol (PubChem CID: 5280863) Isoquercetin (PubChem CID: 5280804) Myricitrin (PubChem CID: 5281673) Oleanolic acid (PubChem CID: 10494) Ursolic acid (PubChem CID: 64945)

Abbreviations: ACAT, acyl coenzyme a-cholesterol acyltransferase; AGEs, advanced glycation endproducts; AHA, alpha hydroxy acids; Ang, angiotensin; AOPPs, advanced oxidation protein products; APTT, activated partial thromboplastin time; ASK 1, apoptosis signal regulating kinase 1; AST, aspartic transaminase; BP, blood pressure; CI, cardiac index; CK, creatine kinase; CO, cardiac output; CPIP, common peak isovolumetric intra-ventricular pressure; CVP, central venous pressure; DHU, 19, 24-dihydroxy ursolic acid; DOX, doxorubicin; ET, endothefin; GSH-Px, glutathione peroxidase; GVH, graft-versus-host; HDL, high density lipoprotein; HK, hexokinase; HR, heart rate; I/R, ischemic reperfusion; LDH, lactate dehydrogenase; LDL, low density lipoprotein; LVEDP, left ventricular end-diastolic pressure; LVP, left ventricular pressure; LVSP, left ventricular systolic pressure; LVWI, left ventricular work index; MACO, middle cerebral artery occlusion; MAP, mean arterial pressure; MBC, minimum bacterial concentrations; MDA, malondialdehyde; MIC, minimum inhibition concentration; 2-NBDG, 2-[N-(7-nitrobenz-2-oxa- 1,3-diazol-4-yl) amino]-2-deoxy-D-glucose; NO, nitric oxide; NOS, nitric oxide synthase; OA, oleanolic acid; PA, proanthocyanidin; PAf, proanthocyanidin fraction; PLE, persimmon leaf extract; PLEg, persimmon leaf extract 20 0 galloly moiety; PLF, persimmon leaves flavonoids; PT, prothrombin time; RA, rotungenic acid; SCF, sinus coronary flow; SOD, superoxide dismutase; SRBC, sheep red blood cell; STZ, streptozotocin; TC, total cholesterol; TCM, traditional Chinese medicine; TG, triglyceride; TPVR, total peripheral vascular resistance; TT, thrombin time; TTI, tension time index; UA, ursolic acid; VCMCs, vascular smooth muscle cells; 4-VO, 4-vessel occlusion n Corresponding author. Postal address: The School of Pharmaceutical Sciences, Sun Yat-sen University, 132 Waihuan Rd. East, Guangzhou Higher Education Mega Center, Guangzhou 510006, PR China. Tel.: þ86 13924061779, þ 86 20 39943041; fax: þ 86 20 39943000. E-mail address: [email protected] (X. Xu). http://dx.doi.org/10.1016/j.jep.2015.01.007 0378-8741/& 2015 Published by Elsevier Ireland Ltd.

Please cite this article as: Xie, C., et al., Persimmon (Diospyros kaki L.) leaves: A review on traditional uses, phytochemistry and pharmacological properties. Journal of Ethnopharmacology (2015), http://dx.doi.org/10.1016/j.jep.2015.01.007i

67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88

C. Xie et al. / Journal of Ethnopharmacology ∎ (∎∎∎∎) ∎∎∎–∎∎∎

2

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 Q3 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66

Contents 1. 2.

Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 Botany and ethnopharmacology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 2.1. Botany . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 2.2. Ethnopharmacology. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 3. Phytochemistry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 3.1. Flavonoids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 3.2. Terpenoids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 3.3. Other compounds . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 4. Analytical methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 5. Pharmacological properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 5.1. Effect on cardiovascular system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 5.2. Antioxidant activity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 5.3. Antiatheroscloresis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 5.4. Antidiabetic activity. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 5.5. Hemostasis activity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 5.6. Anti-anaphylactic activity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 5.7. Antibacterial activity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 5.8. Anticancer and antitumor effect . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 5.9. Immunologic activity. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 5.10. Tyrosinase inhibitory effects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 6. Toxicity studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 7. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 Uncited references . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11 Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11

1. Introduction

2. Botany and ethnopharmacology

“Shi Ye” is the fresh or dry leaves of persimmon (Diospyros kaki L.), which is a kind of plant native to China and widely distributed in tropics and subtropics of East Asia such as Japan and Korea (Mallavadhani et al., 1998). The persimmon leaves were traditional Chinese medicine (TCM), first recorded in Diannan Bencao written by Lan Mao in Ming Dynasty (Lan, 1975): The frosted leaves can cure chronic ulcer. They have been long used in the treatment of stroke and the syndrome of apoplexy in clinic to improve ischemia stroke, angina and internal hemorrhage, as well as paralysis, frostbite, burns, hemorrhage and constipation. It also showed radical scavenging, neuroprotection, thrombosis inhibition, atherosclerosis, anti-mutagenic, antihistamine and anti-allergic properties (Kotani et al., 2000; Matsumoto xet al., 2002; Tanaka et al., 2003; Sakanaka et al., 2005). Besides, they are utilized as a hypotensive drug in Japanese traditional medicine (Funayama and Hikino, 1979). Naoxinqing tablet made from the extract of persimmon leaves have been recorded in the supplement of China Pharmacopeia 2010 as a protected varieties of traditional Chinese medicines for the prevention and cure of the coronary heart disease and cerebral arterial sclerosis. Over the past few decades, there has been a lot of information available on the phytochemistry and pharmacological activities of constituents isolated from persimmon leaves. However, its beneficial outcomes on human health are still unclear due to limited availability of well-designed clinical trials. Indeed, most of the publications are in Chinese literature and recorded base on the TCM theory, which, at time, restricts access to non-Chinese readers. The current review aims to compile an up-to-date and comprehensive literature analysis of persimmon leaves that covers ancient and modern medical applications, phytochemistry, in vitro pharmacological and in vivo pre-clinical studies, toxicology and analytical methods based on information obtained from both English and Chinese literature.

2.1. Botany The family Ebenaceae (about 500 species) is widely spread in tropics and subtropics. The genus, Diospyros (Syn: Persimmon, ebony) with more than 249 species is the most important, both numerically and economically. Among them, persimmon (Diospyros kaki), which originated from China, has been long and widely used as a traditional medicine. It was also reported to be the best fruit yielding species (Mallavadhani et al., 1998). Plant morphology of Diospyros kaki was shown in Table 1 and Fig. 1.

2.2. Ethnopharmacology Persimmon was widely distributed in China, India, Japan, and Korea (Funayama and Hikino, 1979). With a wide spectrum of biological and pharmacological properties, persimmon leaves were traditionally utilized as a medicine, health beverage, and cosmetic. The traditional medical use of persimmon leaves have been recorded in Chinese ancient medical books. Diannan Bencao, which was written by Lan Mao in 1556 in Ming Dynasty, first recorded persimmon leaves as medicine for the treatment of chronic ulcer of the lower part of legs in folk (Lan, 1975). In 1841 (Qing dynasty), the book of Chinese materia medica named Bencao Zaixin (Ye, 1933), described that persimmon leaves could be applied to treat cough, hematemesis, increasing salivation and slaking thirst. Afterwards, according to Fen Lei Cao Yao Xing, a medical book written in the late Qing Dynasty, persimmon leaves could cure lung distension (“Fei zhang” in Chinese traditional theory) (Anons, 1911). In Chinese herbal medicines Guangxi standard (1990s), persimmon leaves were reported to be effective on cough, internal hemorrhage, hypertension, cerebral arteriosclerosis and coronary disease. Besides, they were also used


67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 Q10 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66

3

Table 1 Plant morphology of Diospyros kaki (Xiong et al., 1993). Items

Description

Tree

Tall deciduous trees, reaching 14 m in height. Dark gray barks with rectangular dehiscence and patulous branches with dark brown lenticels (there are pubescence on twigs). Leaves Simple, alternate, ovate elliptic and bovate; 5–18 cm long and 2.8–9 cm wide; Apex acuminate or obtuse with entire cuneate base. Pubescence growing in dark green primary veins, short pubescence growing in light green leaves. Petioles Round, 8–20 mm long. Flowers Blossoms in May. Hermaphrodite. The cyme belongs to male flowers and female flowers are solitary in distal leaf axil. Peduncles: 5 mm long with tiny bract. Lower part of calyx is short cylindrical, cracking to four parts with fur growing inside. Corolla is campaniform and yellow white, craking to four parts. 16 Stamens can be seen in a male flower while 8–16 in a bisexual flower. A single female flower possesses 8 staminodiums, ovary superior with 8 room, style isolating from base of branch. Fruit Fruit in September and October. The pepos are orange yellow and sepal in base. Obovoid or vary in other shapes with the diameter differ from 3.5 to 8 cm. Seeds Brown and oval.

Fig. 1. Persimmon (Diospyros kaki) leaves: A. young leaves; B. typical leaves; C. yellow leaves; D. dried leaves.

traditionally to promote maternal health (Han et al., 2002). At present, Naoxinqing tablet made from the extract of persimmon leaves have been recorded in Chinese Pharmacopeia 2010, and it is a patented and approved drug of Traditional Chinese Medicine (TCM) used for the treatment of cerebral arteriosclerosis (Chinese Pharmacopoeia Commission, 2012). Persimmon leaves have also been applied as the material of health beverages for centuries. The favorite, centuried and widely used preparation is persimmon leaves tea, which is popular with Chinese and Japanese since it helps reduce blood pressure, eliminate cholesterol accumulating, and prevent melanin accumulating. In China, Korea and Japan, Kaki-No-Ha Cha (persimmon leaf tea) has been consumed as a subtle, popular beverage. The persimmon leaves are infused with hot (rather than boiling) water and drunk as kakinoha-cha in the same way as green tea (Sakanaka et al., 2005). In Japan, it has been widely regarded as a famous nutrient and anti-aging tea for the rich vitamin C in persimmon leaves, which makes it great in sales (Mizuo, 1995; Hiromichi, 2002; Tsurunaga et al., 2011). Another finding on

persimmon leaves is that the flavonoids in persimmon leaf tea would be metabolized by intestinal bacteria when brought into alimentary tract, and then absorbed from the gastrointestinal tract into the blood (Bei et al., 2009; Zhang et al., 2014). Moreover, persimmon tea has been proved to be valuable in content of vitamin C, substituting at least in part for the expensive citrus fruits. This tea has a very pleasant flavor, which makes people feel better. Tea made from the green leaves is very acceptable but that from the dried leaves is even better, having a flavor slightly reminiscent of sassafras (Gibbons, 1962). Persimmon leaves were more and more popular not only on the medicinal use but also in the cosmetic and food industries due to its skin-lightening effects (Hanamura et al., 2008). Diannan Bencao has recorded that drinking persimmon leaves in daytime and wash face or body with persimmon leaves decoction would effectively made skin whitener (Lan, 1975). Accordingly, people in Taiwan call it “beauty leaves”, which maybe potentially applied on a skin care product in future. Persimmon leaves were also employed as a natural food additive for healthcare in East Asia.


67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 Q4126 127 128 129 130 131 132


4

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66

It is interesting to notify that the leaves extract of Japanese persimmon in combination with jasmine has been used for making anti-tobacco smoking candies in Japan (Mallavadhani et al., 1998).

3. Phytochemistry 3.1. Flavonoids Flavonoids were the main constituents in persimmon leaves and considered as the active compounds (Liu et al., 2012). The structures of different flavonoids found in persimmon leaves were summarized in Fig. 2, Table 2. Kaempferol, quercetin and the derived glycosides on the 30 -glycosyl, etc. were separated (Cai and Yang, 2001). Chrysontemin isolated from persimmon leaves showed moderate inhibitory activity for tyrosinase, and there are reports that chrysontemin inhibits tumor promoter-induced carcinogenesis and tumor metastasis in vivo. The antifungal activity of hyperoside and trifolin; the anti-inflammatory activity of isoquercitrin; the antiallergic activity of astragalin; and the angiotensin-converting enzyme inhibitory activity of astragalin, isoquercitrin was also reported (Xue et al., 2011). Vomifoliol 9-O-a-arabinofuranosyl(1-6)-β-D-glucopyrano-side from persimmon leaves could augment peripheral glucose as an insulin-sensitizing agent against Type 2 diabetes mellitus (Wang et al., 2011).

Table 2 The main flavonoids in the persimmon (Diospyros kaki) leaves. Compounds Flavonol Quercetin, Kaempferol Isoquercetin Myricitrin Flavonol glucoside Rutin Quereetin-3-O-β-L-arabinopyranoside Quereetin-3-O-β-D-glucopyranoside Quereetin-3-O-β-D-galactopyranoside Kaempferol-3-O-α-L-rhamnopyranoside Kaempfetol-3-O-β-D-galactopyranoside Kaempferol-3-β-D-xylopyranoside Kaempefrol-3-O–L-arabinopyranoside Kaempferol-3-O-(20 0 -O-galloyl)-β-D-glucopyranoside Myrieetin-3-O-α-D-glucopyranoside Quereetin-3-O-β-D-galaetoside (Hyperin)

References

Chou (1984) Nakatani et al. (1989) Guo and Dong (1999) Chou (1984)

Chen et al. (2002)

3.2. Terpenoids Triterpenoid compounds (Fig. 3, Table 3) were also important in persimmon leaves and showed certain pharmacological activities. Ursolic acid (UA), 19-hydroxy ursolic acid and 19, 24-dihydroxy ursolic acid (DHU) isolated from persimmon leaves suppressed stimulus-induced superoxide generation and tyrosyl phosphorylation and may have pharmaceutical applications. (Chen et al., 2002) 3.3. Other compounds Vc was another main compound in persimmon leaves. Its content was reported to be 13.9 mg/g among the determined 410 kinds of leaves (Liang et al., 2000). Further research revealed that Vc in fresh leaves was as high as 25.0 mg/g, which was much higher than that in the fruits of jujube, kiwi, lemon, orange, etc. Clark and Smith demonstrated that the content of Vc and trace elements changed with the seasons, revealing positive correlation (P o0.01) and the highest value was in May (Clark and Smith, 1990).

Fig. 3. Chemical structures of oleanolic acid and ursolic acid

Resins, polysaccharides, chlorophyl (Hospital, 1973), carotene, kryptoxanthin, cellulose, hemicelluloses, lignin (Hu et al., 2002), amino acids and trace elements (Zu and Lei, 1989) were also found in persimmon leaves. Other compounds reported in persimmon leaves were listed in Table 4.

4. Analytical methods Analytical methods of persimmon leaves were focused on flavonoids and their derivatives or triterpenoids by HPLC, as summarized in Table 5.

5. Pharmacological properties 5.1. Effect on cardiovascular system

Fig. 2. The chemical structure of flavonoids.

Persimmon leaves were used to manufacture tea for drink in China and Japan, which has been popular with local people for many years. It is because that persimmon leaves and its preparations possess a great many effective activities on cardiovascular system. Several reports have proved that persimmon leaves increased the coronary artery blood flow in the hearts of rabbits and frogs in vitro and the coronary blood circulation in anesthetic dogs (Han et al., 1983; Liang and Fu, 1985). Huang (Huang et al., 1983) evaluated the effects of alcohol extract from persimmon


67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66

5

Fig. 4. HPLC-DAD profiles of flavonol 3-O-glycosides in persimmon leaves during five growth stages in 2007.

Fig. 5. Separation of a standard mixture of RA, OA, and UA by HPLC, flow rate: 1 mL min 1, mobile phase: methanol-0.2% aqueous AcOH (85:15, v/v) in the presence of increasing concentrations of HP-β-CD (10 mmol L 1) at 35 1C. (RA. Rotungenic acid; OA. Oleanic acid; UA. Ursolic acid).

Fig. 6. HPLC fingerprints of 12 batches of persimmon leaves samples.

leaves on several cardiovascular indexes in anesthetic dogs. Partial results were listed in Table 6. However, there were no significant effects on other indexes including LVSP, CVP, dp/dtmax, dp/dt/CPIP, t dp/dtmax and LVWI between 50 mg/kg group and normal group. Moreover, significant differences between the dose of 50 and 100 mg/kg were observed in the increases of CO (P o0.05) and SCF (P o0.01). The above results demonstrated that persimmon leaves could improve

cardiac pumping function but had no significant effects on myocardial contraction, which was not because of its direct effects on myocardium but decreasing TPVR and arterial blood pressure. It was reported that persimmon leaves exhibited protective activities against injuries caused by myocardial ischemia. In Deng's experiment (Deng et al., 2004), the acute myocardial ischemia model was induced by ligating left anterior descending coronary artery at the distal one third segment in rat, open chest rats. The epicardial electrocardiogram was examined before and after ligation, the infarct aera was showed with TTC stain 21 h after ligation. Furthermore, the effect of leaf of persimmon oral solution on the above parameters and CK and LDH contents of serum were also studied in comparison with that of Fufang danshen diwan and Tabellae isosorbidi dinitratis (lente liberantes). Results: leaf of persimmon oral solution markedly acted against the elevation of S-T segment in the electrocardiograms of rats with acute myocardial ischemia caused by ligating left anterior descending coronary artery at the distal one third segment in rat. The groups using large (100 mg/kg) and small dosage (25 mg/kg) of oral solution made from persimmon leaves markedly reduced the incidence of arrhythmia and the area of myocardial infarction, and lowered the CK and LDH contents of serum. Furthermore, persimmon leaves flavonoids of (PLF) at the dosage of 4 mg/mL could reduce the releasing of AST, CK and LDH and producing of MDA, as well as the increasing activities of SOD, Na þ –K þ ATP enzyme and ATP-Ca2 þ enzyme, which revealed that PLF had the myocardial protective effects against I/R injury on rats (Sun et al., 2009). Similar effect could be implied by Bei's experiment (Bei et al., 2009), shown in Tables 7 and 8. FLDK-P70 (a standardized flavonoids extract from persimmon) significantly protected rats from MCAO and 4-VO ischemic injury in vivo and protected hippocampal neurons from glutamate-induced excitotoxic injury as well as cortical neurons from hypoxia-induced injury in vitro. The mechanisms of these effects might be due to the antioxidative activity of the flavonoids, implying that FLDK-P70 might be useful for the prevention and treatment of ischemia/reperfusion injury and other related neurodegenerative diseases.


67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132


6

1 2 3 4 5 6 7 8 9 10 11 12 Q8 Q9 13 Q7 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66

Table 3 The main terpenoids in the persimmon leaves. Name

Group

Oleanolic acid α-Amyrin, Ursolic acid Uvaol, Uvasol, 19α-Hydroxyursolie acid, 19α, 24Dihydroxyursolic Lupeol, Betulinic acid, Betulic acid

Oleanane Ursane

Pomolic acid

References

Tezuka et al. (1972) Tezuka et al. (1972), Matsura and Linuma (1971), Chen et al. (2000) Lupane Tezuka et al. (1972), Zakaria et al. (1984), Matsura and Linuma (1971) Chrysobalanaceae Thuong et al. (2008)

Table 4 Compounds reported in persimmon leaves. Compounds

Phyto-constituents

References

Naphthoquinones

3-Bromo-plumbagin 3-methoxy-7-methyl-uglone, Martinone, 8'-hydroxy-isodiospyrin Diospyrin, Isodiospyrin, Neodiospyrin Mamegakinone, 7-Methyl-uglone

Organic acid

Succinic acid, Benzoic acid, Salicylic, Pyromucic acid, Syringic acid, Hydroxybenzoic acid, Procatechuic acid, Indoleacetic acid

Tezuka et al. (1972), Zhong et al. (1984) Lin et al., (1988) Tezuka et al. (1972), Lin et al. (1988) Sidhu et al. (1968), Tezuka et al. (1972), Zhong et al. (1984), Lin et al. (1988) Zhou et al. (1987)

Anionic

H2 PO4 , NO3 , SO4 2 , Cl Campesterol, stigmasterol, β-sitosterol 6–7-hydroxyl-7-hydroxy coumarin, scopoletin Myristic acid, palmitic acid, stearic acid, 10-octadecenoic acid, cerotic acid linoleic acid, linolenic acid,.etc. Shibuol, Colorless delphinium meal-3-glycosidase

Phytosterol Coumarin Flavor substances and fatty acids Tannins

Zhang et al. (2002) (Behan et al., 1996) Zhou et al. (1987) An and Guo (2000). Medicine (1978)

Table 5 HPLC determination of constituents in persimmon leaves. Analytes

Column

Mobile phases

Persimmon leaf flavonols

Inertsil ODS-3 column (250 4.6 mm i.d., 4 μm; GL Science Inc., Tokyo, Japan) Agilent Zorbax SB-C18 reserved phase column (5 μm, 250 mm 4.6 mm, i.d.) Kromasil 5Uc C18 column (100A 250 4.6 mm)

(A) 0.5% v/v phosphoric acid 350 nm in water and (B) 0.5% v/v phosphoric acid in acetonitrile Methanol–0.1% aqueous 210 nm H3PO4 (88:12, v/v)

barbinervic acid (BA) rotungenic acid (RA), 24-hydroxy ursolic acid (HA) Rutin and oleanolic acid

Wavelength

Quercetin and kaempfetol

Methanol–water (55:45, v/v) for rutin; Acetonitrile– methanol–water (70:16:14, v/v) Hypersil C18 column Methanol–0.4% aqueous (250 mm 4.6 mm, 5 μm) H3PO4 (50:50, v/v)

365 nm rutin and 215 nm (oleanolic acid) 360 nm

Oeanolic acid and ursolic acid

Kromasil C18 column (5 μm, 250 4.6 mm)

Methanol–0.2% aqueous H3PO4 (87:13, v/v)

210 nm

Fingerprint

phenomenex luna C18 column (5 μm, 250 4.6 mm)

Acetonitrile-10 mmol/L monopotassium phosphate (gradient elution)

360 nm

Persimmon leaf flavonoid (PLF) markedly reduced the blood pressure (BP) and heart rate in normal rats. After intragastric administration of PLF at the dose of 20, 40 and 80 mg/(kg d) for 4 weeks, BP significantly reduced with a dose-dependent manner, as compared with normal control group. PLF could increase the serum NO (77.15715.14 vs 30.77710.62 μmol/L, Po0.01), and decrease the plasma ET (56.2179.23 vs 87.28711.75 pg/mL, Po0.01) and Ang II (503.4733.40 vs 666.62733.45 pg/mL, Po0.01) as compared with model group (distilled water). It is suggested that PLF may adjust the imbalance of cardiovascular active substances by increasing the release of endogenous vasodilators and reducing the release of endogenous vasoconstrictors, resulting in anti-hypertension (Qin et al., 2008). Water-soluble proanthocyanidin fraction, the major polyphenols in persimmon leaf tea, was found to significantly decrease blood pressure after 4 h since oral administration of 300 mg/kg in

Results

Reference

4 20 0 -galloylated flavonol glycosides had accumulated in developing leaves of persimmon by the end of May through a rapid 20 0 -galloylation of the corresponding non-galloylated flavonol glycosides (Fig. 4) BA, RA and HA were 0.46–1.43%, 0.13–0.68% and 0.09– 0.28% in persimmon leaves, respectively. (Fig. 5)

Kawakami et al. (2011)

The leaves of persimmon collected in July and October have the higher contents of total flavonoids rutin and oleanolic acid.

Yuan et al. (2006)

The contents of quercetin and kaempfetol were 3.58– 3.88% and 6.18–6.42% in the extract of persimmon leaves, respectively. The contents of oleanolic acid and ursolic acid were 0.05–0.17% and 0.24–0.59% in the extract of persimmon leaves, respectively. Similarity values of 12 batches of samples from Shandong, Shanxi and Anhui province, China were 0.78– 0.99 (Fig. 6)

Bei et al. (2005)

Fan et al. (2011)

Zou and Sun (2009) Liu et al. (2008)

SHRs was performed. However, no change was observed in normotensive WKY rats. The proanthocyanidins levels of endothelial nitric oxide synthase (Ser-1177) and the upstream kinase Akt (Ser-473) in umbilical cells also increased in a time-dependent manner after the addition of a proanthocyanidins, which was due to vasorelaxation via an endothelium-dependent nitric oxide/cGMP pathway, and that proanthocyanidin might be useful in dietary lowering of blood pressure (Kawakami et al., 2011a). Persimmon leaves in treatment of cardiovascular diseases were mainly attributing to inhibitng the myocardial apoptosis, which was the basis of cardiovascular disease. Advanced Glycation Endproducts (AGEs) was an important factor in the process of apoptosis. They were believed to induce changes in endothelial cell properties relevant in the pathogenesis of vascular disease(Bierhaus et al., 1998). In Ouyang's experiment, apoptosis of neonatal rat cardiac myocytes was induced


67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66

by hypoxia-reoxygenation in a time-dependent manner. The flavone extracted from persimmon leaves can inhibit apoptosis of cultured neonatal rat cardiac myocytes induced by hypoxia-reoxygenation and AGEs in vitro (Ouyang et al., 2003a). The flavones of persimmon leaves used for treatment of the cells significantly inhibit the proliferation of adventitial fibroblasts stimulated by AGEs in vitro (Ouyang et al., 2003b). Persimmon leaves have also been proved to possess the activity in curing vascular occlusive diseases (e.g. atherosclerosis) and the mechanism was related to fibroblasts, which were the main cells of arterial blood vessels. Advanced oxidation protein products (AOPPs), structurally similar to AGEs-proteins and exerting similar biological activities as AGEs, i.e. induction of proinflammatory cytokines and adhesive molecules, could significantly induced fibroblasts proliferation in the adventitia. Chen (Chen et al., 2002) found that five triterpenoid compounds isolated from persimmon leaves had obvious effect on stimulus-induced superoxide generation and tyrosyl phosphorylation in human polymorphonuclear leukocytes, assuming that it inhibited fibroblasts proliferation induced by AGEs and AOPPs. In addition, VSMCs proliferation and ASK1 expression induced by TNF (20 ng/mL) in 6-week old male SD rats were significantly inhibited by PLF (50 μg/mL) using MTS/PES method (P o0.05) (Ouyang et al., 2007). The above results were really a hint that the persimmon leaves could be developed as a leading drug in curing cardiovascular disease. 5.2. Antioxidant activity Persimmon leaves also exerted potential antioxidant activity in vitro. Ethanolic extracts from persimmon leaves (600 mg/kg) was effective in retarding lard deterioration, which showed a little higher antioxidative activity than hesperidin (100 mg/kg) and tea polyphenol (100 mg/kg) (Shi et al., 1999). Total flavonoids from persimmon leaves significantly decreased the level of reactive oxygen species and malondialdehyde, with the increasing activity of catalase, SOD and glutathione peroxidase (GSH-Px) in E1 cells in a dose-dependent manner (Sun et al., 2011), indicating that persimmon leaves extract and persimmon leaf tea exerted the

Table 6 The effects of alcohol extract from persimmon leaves on several cardiovascular indexes in anesthetic dogs. Index

Medicated group 50 mg/kg 100 mg/kg

Normal control group

SCF mAPHR TTIHR TPVR ETHR CI

↑a ↓a ↓a ↓a ↓a –b

– – – – – –

↑a ↓a ↓a ↓a ↓a ↑a

“↑”-increase, “↓”-decrease, “–”-no effect. a b

P o0.05. P40.05.

effect of clearing oxygen free radical and antioxidant. To investigate their mechanisms in osteoblast cells injured by oxidative stress, Sun et al. (2014) addressed the potential therapeutic or toxic effects on H2O2-stimulated MC3T3-E1 cells and has found that flavonoids from persimmon leaves (FPL) attenuated H2O2induced apoptosis in MC3T3-E1 cells via the NF-kB pathway. The results suggest that the molecular mechanism of FPL in antiapoptosis was associated with the suppression of the translocation of NF-kB/p65 into the nucleus. The protective effect of FPL could provide a promising approach for the treatment of osteoporosis. Several studies have proved that the persimmon leaves possess free radical scavenging activity (Han et al., 2002; Sakanaka et al., 2005). Han's trial showed the IC50 value of methanol extract of persimmon leaves was 0.11 mg/mL against DPPH radical (Han et al., 2002). The effect of PLF in free radical scavenging of DPPH was not as good as rutin, as EC50 were 96.36 72.63 and 41.56 71.96 μg/mL, respectively (Sun et al., 2011). PLF possessed scavenging activity of superoxide anion (EC50, 41.58 70.21 μg/mL) and hydroxyl radical (70.647 3.22), reducing power and iron chelating activity, which was significantly better than that of rutin (Sun et al., 2011). The scavenging activity against superoxide anion radicals of different persimmon leaves extract has also been compared. The effect of methanol extract of persimmon leaves was stronger than that of the water extract (Fig. 7). DPPH radical-scavenging activity of water and methanol extracts was strong, and 0.1% water extract showed an inhibition rate of more than 90%. The hydroxyl radical-scavenging activity of the 0.1% extracts was nearly equal to that of 10 mM ascorbic acid (Fig. 8) (Sakanaka et al., 2005). Also, PLF scavenged OH∙ to inhibit the MDA produced by OH∙ from the tissue, hepatic mitochondria and hepatic microsome of rats, so as to reduce erythrocyte hemolysis, swelling of mitochondria and hemolysis of RBC (Yang et al., 2003). It has been demonstrated that pretreatment with FLDK-P70 alleviated H2O2-induced cell injury and apoptosis, by up-regulating Bcl-2 expression and improving redox imbalance, indicating the alleviation of the decline in the intracellular endogenous antioxidants: glutathione and glutathione peroxidase as well as catalase with the decreasing leak of lactate dehydrogenase and the reduction of malondialdehyde accumulation. It implied that FLDK-P70 might be potentially used in the prevention and treatment of ischemia/reperfusion injury and other neurodegenerative disease (Bei et al., 2005). Antioxidant activity of persimmon leaves was not only induced by PLFs but also by triterpenoid compounds. Chen (Chen et al., 2002) reported that when the cells were pre-incubated with five triterpenoid compounds isolated from persimmon leaves, the superoxide generation induced by N-formyl-methionyl-leucyl-phenylalanine was significantly suppressed in a concentration-dependent manner. These compounds also suppressed the superoxide generation induced by arachidonic acid in high concentrations. Although PLF have exerted antioxidative activity in vitro, the function of PLF on humans is still unconvincing because humans were not involved in those studies. The antioxidative activity might be very limited since PLF is admittedly hard to absorb, which leads to low bioavailability. Even so, some flavonoids, such as rutin, can be highly metabolized by intestinal bacteria after oral intake (Zhang et al., 2014).

Table 7 The myocardial protective effects of FLDK-P70n(4-VO (four-vessel occlusion) model & MCAO (middle cerebral artery occlusion)). Dosage 1. 40, 80 mg/kg body weight; p.o. 2. 40, 80 mg/kg body weight; p.o.

Administration time

7

Observation

5 days before and 7 days after 4-VO

Increased the survival of hippocampal CA1 pyramidal neurons after transient global brain ischemia in rats. 3 days before and 4 days after middle cerebral artery Significantly reduced the lesion of the insulted brain hemisphere while improved the occlusion (MCAO) neurological behavior of rats


67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132


8

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66

Table 8 The myocardial protective effects of FLDK-P70a (Neuronal death and apoptosis).

1. 2. a

Pretreatment concentration

Observation

5, 10 μg/ml 25, 100 mg/ml

Protected neurons from glutamate-induced excitotoxic neuronal death. Reduced hypoxia-reoxygen induced neuronal death and apoptosis in a dose-dependent manner.

FLDK-P70 (a standardized flavonoids extract from persimmon).

Fig. 7. Superoxide anion radical-scavenging activities of water and methanol extracts of persimmon leaf. Ascorbic acid (asc) was used as positive control. (a) Water extract; (b) methanol extract. Values are means 7SD (n¼ 3).

Fig. 8. DPPH radical-scavenging activities of water and methanol extracts of persimmon leaf tea. Ascorbic acid (asc) was used as positive control. (a) Water extract; (b) methanol extract. Values are means 7SD (n¼ 3).

5.3. Antiatheroscloresis Persimmon leaves extract and leaf tea showed anti-atheroscloresis effect by decreasing the triglyceride (TG) and total cholesterol and LDL-C level, whereas increasing the ratio of HDL level in serum of high-fat diet fed SD rats (Batchelor et al., 1987; Lin et al., 1996). 40 wistar's rats were fed to high-fat diet with drinking 1 mL persimmon leaves juice (25%, w/v) for 6 weeks and the body weight level in experimental observation group (1 mL distilled water) were obviously lower than that of experimental control group (Po0.01). In the meantime, the TG, TC and LDL-C levels in serum of experimental observation group were obviously lower and the HDL-C levels were significantly increased (Po0.05), as compared with that of experimental control group (Wu and Wang, 1999). Persimmon leaves diminished the accumulation of hepatic lipid droplets and the epididymal white adipocyte size in high-fat rats but improved hepatic HMG-CoA and ACAT activities. Studies also found that the content of fecal TG, cholesterol and acidic sterol were

significantly higher in high-fat with 5% (wt/wt) powdered persimmon leaves group. The results indicated that persimmon leaves improved plasma and hepatic lipid levels profile, partly via the increased fecal lipids in high-fat fed rats. It might be due to the properties of the phenolic compounds and high fiber content in the powdered persimmon leaves (Lee et al., 2006). In another case, a cholesterol-free diet containing dried powder of persimmon leaves at 5% level of dietary fiber was fed to male SD rats, the fecal weight and fecal excretion of bile acids per gram or per day were significantly increased (Innami et al., 1998). In a brief, Persimmon leaves could be considered to be explored as an anti-atheroscloresis drug, since it exerted prominent anti-atheroscloresis effect. 5.4. Antidiabetic activity The ethanol fraction, ethylacetate fraction, n-butanol fraction and water extract fraction precipitated by alcohol from persimmon leaves significantly decreased blood glucose in STZ-induced


67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 Q5124 125 126 127 128 129 130 131 132


1 2 3 4 5 6 Q6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66

diabetic mice, with the decreasing insulin resistance index and significantly increased insulin sensitivity index (Deng et al., 2011). The significant effect of water extraction was resulted from the polysaccharide of persimmon leaves, which reduced blood glucose level and improved glucose tolerance, with the increasing activity SOD, GSH-Px, HK and decreasing MDA content in liver (Hu et al., 2006). In Jung's study (Jung et al., 2012), powered PL (5%, w/w) was supplemented with a normal diet to C57BL/KsJ-db/db mice for 5 weeks. PL decreased blood glucose, HOMA-IR, plasma triglyceride and total cholesterol levels, as well as liver weight, hepatic lipid droplets, triglycerides and cholesterol The antihyperglycemic effect was linked to decreased activity of gluconeogenic enzymes as well as increased glycogen content, glucokinase activity and its mRNA level in the liver. Also, PL led to a decrease in lipogenic transcriptional factor PPARc as well as gene expression and activity of enzymes involved in lipogenesis, with a simultaneous increase in fecal lipids, which are seemingly attributable to the improved hyperlipidemia and hepatic steatosis and decreased hepatic fatty acid oxidation. As a result, PL may be an effective dietary strategy to improve type 2 diabetes accompanied by dyslipidemia and hepatic steatosis by partly modulating the activity or gene expression of enzymes related to antioxidant, glucose and lipid homeostasis. Another exciting news was that a novel glucosidase inhibitor, vomifoliol 9-O-α-arabinofuranosyl (1-6)-β-D-glucopyranoside was isolated for the first time from persimmon leaves. It exhibited a strong anti-α-glucosidase activity with an IC50 value of 170.62 nM and stimulated a dose-dependent increase in the uptake of a fluorescent D-glucose analog, 2-[N-(7-nitrobenz-2oxa-1,3-diazol-4-yl) amino]-2-deoxy-D-glucose (2-NBDG), in HepG2 cells at a rate higher than that of insulin controls. It was also found to be associated with adipocyte differentiation and moderate increased in 2-NBDG uptake by 3T3-L1 cells. These findings suggested that vomifoliol 9-O-α-arabinofuranosyl (1-6)β-D-glucopyrano-side could augment peripheral glucose as an insulin-sensitizing agent against Type 2 diabetes mellitus (Wang et al., 2011). 5.5. Hemostasis activity Persimmon leaves were traditionally used as a hemostatic. In 1956, Yamashita reported the clinical use of persimmon leaves for the hemostasis activity. Persimmon leaves reduce mucosal bleeding and prevent gastrointestinal ulcers (Funayama and Hikino, 1979). Then He (He et al., 1986) found that content of flavonoids and their derivatives were higher (1.577–1.638%) when analyzing the constituents in Hemostatic No. 4 (a preparation consisted of persimmon leaves powder). In 2005, Wang evaluated the hemostasis activity of isoquercitrin and astragalin isolated from persimmon leaves. Oral administration to 10 mice per group at 10 mg/kg was conducted to determine bleeding time. Compared with normal group (7.277 1.33 min), the bleeding time was both shorter in isoquercitrin (6.98 71.36 min) and astragalin group (5.62 71.40 min) (P o0.05). Beside the flavones from persimmon leaves, other fractions were also studied for the hemostasis activity. Sa (Sa et al., 2005) found the purified fraction from persimmon leaves had antithrombotic activity since it delayed thrombin time (TT), activated partial thromboplastin time (APTT), and prothrombin time (PT) using human plasma. TT was prolonged 2 and 3 times more than the control value at 5.3 and 13.1 pg/mL of the anticoagulant from persimmon leaves, respectively. Compared to its effects on TT, it delayed APTT and PT to a lesser degree. APTT and PT required 27.0 and 30.0 μg/mL of the anticoagulant to reach double the control value, respectively. The results suggested that the anticoagulant activity might be caused by degradation or a defect of fibrin or

9

thrombin. It did not cause the hydrolysis of fibrin after incubation but inhibited thrombin-catalyzed fibrin formation with a competitive inhibition pattern. Persimmon leaves also had the hemostatic function on metrorrhagia by drug abortion of pregnant rats in early and middle stage (Wang et al., 2004). In the study of the mechanism of persimmon leaves in curing drug abortion, the Endothelin (ET) content in uterus homogenate of rats from persimmon leaves group was obviously higher than that in blank control group, as well as the content of NO and NOS (nitric oxide synthase) with the significance difference (Po0.01). The ET/ NO in large dosage persimmon leaves group (20 g/(kg d) for 5 d) was obviously higher than that in blank control group and mifepristone-induced abortion control group (3.0 g/(kg d) for 3 d) with the significance difference (P o0.05). The mechanism of persimmon leaves on uterine haemorrhagia after medicine induced abortion might be related with accommodating the balance of NO and ET contents in uterus tissue (Yu et al., 2005). 5.6. Anti-anaphylactic activity It has been reported that persimmon leaves extract or astragalin inhibited histamine release from human basophilic cell line KU812 in response to cross-linkage of FcεRI. Oral administration of both substances dose dependently inhibited passive cutaneous reactions. Moreover, oral administration of these substances to NC/Nga atopic dermatitis-model mice led to a striking suppression of the development of dermatitis, scratching behavior, and serum IgE elevation (Kotani et al., 2000). Oral administration of persimmon leaves extract (250 mg kg 1) or astragalin (1.5 mg kg 1) injection for 4 weeks into NC/Nga mice with overt dermatitis resulted in a decrease in the severity of the condition. The preventive effect on the dermatitis was dosedependent and continuous intake significantly decreased its onset and development. In addition, transepidermal water loss was also suppressed at a persimmon leaf extract dose of 250 mg kg 1. No significant adverse reaction by these substances could be observed (Matsumoto et al., 2002). 5.7. Antibacterial activity The MIC (minimum inhibition concentration) and the MBC (minimum bactericidal concentrations) of persimmon leaves were determined as 0.313 and 0.625 g/mL against tested food spoilage and food-borne pathogens (Escherichia coli, Staphylococcus aureus, Fluorescence pseudomonas, Bacillus subtilis, Bacillus cereus, Proteus vulgaris) respectively, and the antimicrobial rate was over 90%. Furthermore, the activity compounds had been isolated and elucidated as volatile oil, total flavonoid, coumarins and organic acids (Ji et al., 2003). Among those compounds, the volatile oil from persimmon leaves had powerful inhibitory effects against Staphylococcus aureus and Bacillus subtilis, but it had weak inhibitory effects against Escherichia coli. The MIC to Staphylococcus aureus, Bacillus subtilis and Escherichia coli were: 3.13 mL mL 1, 3.13 mL mL 1 and 25.00 mL mL 1, respectively (Chen et al., 2010). Extracts of alcohol and acetic ester from persimmon leaves had remarkable bacteriostasis effects against 6 bacteria. Aqueous extract was more effective against fungi than against bacteria. Petroleum ether extract had no effect against bacteria or fungi. NButanol extract had different effects against bacteria, but had no effects against fungi. Acetic ester extract under different concentrations were tested, which showed that the higher concentration, the stronger bacteriostasis effect was. The sequences of bacteriostasis activities were different under different concentrations (Ji et al., 2006).


67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132

10

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66


Recently, Arakawa et al. (Arakawa et al., 2014) found that persimmon leaves were 6.2 10 5 mol/mL as hydrogen peroxide, and persimmon (Diospyros kaki) leaves extract showed antibacterial activity and inhibition against Escherichia coli (8.7 105 CFU/ mL) and Staphylococcus aureus (2.2 105 CFU/mL), as well as Campylobacter sputorum, Streptococcus mutans, and Bacteroides thetaiotaomicron, which has been demonstrated to be due to the tannins in the plant. 5.8. Anticancer and antitumor effect Persimmon leaves were reported to inhibit the nitrosamineinduced squamous epithelial hyperplasia and cancer in rats (Han et al., 1983). Moon found the anti-tumor effect on sarcoma-180 in vivo (Moon, 1996). Treatment with PLE (persimmon leaf extract) and PLEg (200 galloly moiety) significantly increased the cytotoxicity of doxorubicin (DOX) in A549 adenocarcinoma cells. PLE and PLEg reduced the phosphorylation of checkpoint proteins such as structural maintenance of chromosomes 1, checkpoint kinase 1, and p53 in DOX-treated of ataxia telangiectasia mutated in a dose-dependent manner. PLE, and especially PLEg, abrogated the G2/M checkpoint during DOX-induced DNA damage. These results suggested that PLEg might be a useful sensitizer in cancer chemotherapy (Kawakami et al., 2011b). 5.9. Immunologic activity The effect on immunologic system of Dk-N3 (extract of persimmon leaves) was investigated. Dk-N3 (150, 300 mg/kg, ip, qd 7d) had no significant impact on the weight of spleen and thymus of CFW mice (14–16 g). HC50 caused by giving 150, 300 mg/kg of Dk-N3 and cyclophosphamide (20 mg/kg) to 18– 22 g CFW mice was significantly lower than that of control group (P o0.05), with the sensitization by 3 102 SRBC ip, suggesting Dk-N3 significantly inhibited the formation of antibody. The effect of Dk-N3 on transformation of lymphocyte from cardiac blood was evaluated by 3H-TDR in vitro, using ConA 10 μg/ml as inducing reagent. The transformation was significantly inhibited when DkN3 was more than 25 μg/ml (P o0.01). In addition, when comparing the effect on spleen index of DBA mice with control group, 300 mg/kg of Dk-N3 was significantly lower (Po0.01), indicating that Dk-N3 exhibited significant inhibiting effect on GVH. (Zeng et al., 1987). The inhibitory activity of Dk-N3 on humoral immunity might result in reducing harmful influences on cardiovascular tissue by inhibiting the producing of antibody or improve inflammatory cardiovascular risk indexes. 5.10. Tyrosinase inhibitory effects Triterpenoids with a 3β-hydroxy group were found to inhibit protein tyrosine phosphatase 1B activity, with IC50 values ranging from 3.1 70.2 to 18.8 71.3 mM, whereas those with a 3α-hydroxy moiety were not active (Thuong et al., 2008). Hyperoside, isoquercitrin, trifolin, astragalin, chrysontemin, quercetin-3-O-(200 -O-galloyl-β-D-glucopyranoside), and kaempferol-3-O(2-O-galloyl-β-D-gluco-pyranoside) were isolated from the leaves of six selected persimmon cultivars. Their inhibitory activity was tested against tyrosinase for the oxidation of L-DOPA, and only chrysontemin showed inhibitory activity with IC50 ¼21172 μM), while the other polyphenols, hyperoside, isoquercitrin, quercetin-3-O-(200 -O-galloyl-βD-glucopyranoside) (QOG), trifolin, astragalin, and kaempferol-3-O(200 -O-galloyl-β-D-glucopyranoside) (KOG), did not exhibit any inhibitory activity against the L-DOPA oxidation up to 1 mg/mL (41.6 mM). The tyrosinase activity was effectively reduced by their aglycons, cyanidin (IC50 ¼9.0770.10 μM) and quercetin (IC50 ¼ 9.6770.30 μM)

and, to a lesser extent, also by kaempferol (IC50 ¼50.170.3 μM). It was confirmed that the most influential moiety for tyrosinase inhibition was the 30 , 40 -dihydroxy groups of the catechol moiety (Xue et al., 2011).

6. Toxicity studies Toxicity cases were not found in various uses of persimmon leaves during past hundred years. Modern toxicity studies on animals did not show toxicity in the leaves. In acute toxicity test, after pretreatment with water extract of persimmon leaves, LD50 in male and female mice were both higher than 21.5 g/kg (equal to 597.2 g/kg in raw medical material), suggesting that the extract was of non-toxic grade. In mouse bone marrow micronucleus test (MNT), the ratio of polychromatic erythrocytes/normorchromatic erythrocytes (PCE/NCE) fall in normal range at 10 g/kg (water extract), as compared with that of 20 mg/kg of cyclophosphamide, which implied that persimmon leaves extracts had no mutagenic effect on somatic cells. Also, 10.0 g/kg of persimmon leaves extracts did not show effect of sperm malformation (Wu et al., 2012). In Chen's study (Chen et al., 2005), subchronic toxicity test was performed on 100 SD rats with oral administration of 0.5, 1.0, 3.0, 6.4 g/kg preparations (ethanol extract of persimmon leaves) for 90 days. The blood and physiologic indexes of the rats were not significantly different from that of the normal diet control group. Teratogenicity was evaluated using 100 pregnant rats at the doses of 1.0, 3.0, 6.4 g/kg of preparations, and was compared with negative and positive control group. Significant changes in weight gains were not observed at each dose. The average amount of live fetus, absorbing fetus and dead fetus were not significantly different with negative control group. Anomaly of physiological traits was also not observed at all of doses. Thus, NOAEL of ethanol extract from persimmon leaves was 6.4 g/kg, at the dose of which maternal toxicity, embryotoxicity and teratogenesis were not observed. The long-term toxicity of Naoxinqing Tablets (persimmon leaves extract) has been examined by the experimental study on rats. The rats were intragastric administration Naoxinqing tablets at the doses of 35, 70, 140 g/kg once a day for 180 days. The food intake of the high dose group (140 g/kg) had a slight decrease, the serum total bilirubin had slightly falling for the medium dose group (140 g/kg) and high dose group (140 g/kg) comparing with the control group; no abnormal changes were observed for other indicators. The safe dose for persimmon leaves extract was 35 g/kg (Cai et al., 2012). Now above available toxicity studies showed no toxic activities in persimmon leaves, which implied a reliable safety in common use. However, further studies on toxicity were quite considerable and necessary, especially in different effective extracts, to fill in the blank nowadays.

7. Conclusions Herein, we documented the existing phytochemistry, pharmacological properties and application researches on persimmon leaves. The amount of experimental data evidenced rich nutrients and vast biological active substances in persimmon leaves. A peruse of available scientific references show that the traditional medical uses of persimmon leaves have been evaluated by modern pharmacological studies. Persimmon leaves have the potential multiple pharmacological and therapeutic activities including cardio-protective, antioxidant, anti-inflammatory, anti-atherogenic, antidiabetic, hemostasis, etc., which can be explained by


67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132


1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 Q2 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66

the presence of various flavonoids and terpenoids in the herb. Most of the mentioned pharmacological studies have provided some suggestive scientific evidence for its various traditional usages in ischemia stroke, angina and internal hemorrhage, as well as paralysis, frostbite, burns, hemostasis, constipation, microbial inhibition in Asian countries, especially in China, Korea and Japan. More importantly, there have been no side effects or toxicity reports from many years of research on this herb. In a word, persimmon leaves have received much interest. However, future studies are necessary to address issues regarding composition of the extract, explicability of preclinical experiments and lack of transformation of the preclinical results to clinical efficacy. As a result, persimmon leaves were still employed as folk prescription and the related health products are unpersuasive. Thereby, it is extremely important to conduct detailed investigations on the composition and pharmacological significance of medicinal plants and standardize the formulations based on ingredients. Further systematic studies are necessary to evaluate the efficacy using standardized extracts of persimmon leaves, and to identify the bioactive molecules responsible for the biological activities so that cost-effective, potential medicinal drug and health products can be developed at a large scale. Also, attempts should be made to conduct serious randomized human trials and determine modes or mechanisms of action, bioavailability, pharmacokinetics and physiological pathways for specific bioactives of persimmon leaves which might be responsible behind the protective effects offered by extracts rich in flavonoids and terpenoids in many pharmacological studies. As more scientific evidences on therapeutic effects of Diospyros kaki were found, products (e.g. health care products) based on it might boom in the future.

Uncited references Yu et al. (1988), Fan and He (2006).

Acknowledgments This work was supported by the Bidding Project of GuangdongHongkong breakthroughs in key areas (No. 2010A032100004) and the Industry-University-Research Cooperation Program from Science and Technology Department of Guangdong Province (2013B090200059). References An, Q.R., Guo, Z.F., 2000. Analysis of fatty acids in persimmon leaves by GC–MS. Journal of Instrumental Analysis 19, 74–76. Anons, 1911. Fenlei Caoyao Xing. Chengdu Chengde Tang Version, Chengdu. Arakawa, H., Takasaki, M., Tajima, N., Fukamachi, H., Igarashi, T., 2014. Antibacterial activities of persimmon extracts relate with their hydrogen peroxide concentration. Biological and Pharmaceutical Bulletin advpub 37, 1119–1123. Batchelor, D.B., Carter, M.D., Chen, G.L., Hedrick, C.L., 1987. Coupled model of wave damping, quasilinear heating, and radial transport applied to bumpy tori. Physical Review Letters 58, 2664–2667. Bei, W.J., Luo, J., Wu, A.L., Peng, W.L., 2005. Determination of quercetin and kaempferol in Diospyros kaki leaf extract by HPLC. Chinese Traditional and Herbal Drugs 36, 1014–1015. Bei, W.J., Zang, L.Q., Guo, J., Peng, W.L., Xu, A.L., Good, D.A., Hu, Y.M., Wu, W., Hu, D.H., Zhu, X.H., Wei, M., Li, C.Y., 2009. Neuroprotective effects of a standardized flavonoid extract from Diospyros kaki leaves. Journal of Ethnopharmacology 126, 134–142. Bierhaus, A., Hofmann, M.A., Ziegler, R., Nawroth, P.P., 1998. AGEs and their interaction with AGE-receptors in vascular disease and diabetes mellitus. I. The AGE concept. Cardiovascular Research 37, 586–600. Cai, Y.D., Yang, S.F., 2001. Effect of Nao Xin Qing tablet for cerebral atherosclerosis and angina pectoris of coronary heart disease: an observation of 60 cases. Traditional Chinese Drug Research & Clinical Pharmacology 12, 414–416. Cai, Y.M., Xiang, Z.S., Song, Z.H., Zhang, Z.P., Liu, C.X., 2012. Experimental study on long-term toxicity of Naoxinqing tablets in rats. Chinese Traditional and Herbal Drugs 43, 752–756.

11

Chen, B.F., Huang, J.M., Bei, W.J., Huang, J.K., Bin, T.J., 2005. Study on the subchronic toxicity and teratogenesis of persimmon leaves ethanol extract for 90 day. Toxicology 19, 326–327. Chen, G., Lu, H.W., Wang, C.L., Yamashita, K., Manabe, M., Xu, S.X., Kodama, H., 2002. Effect of five triterpenoid compounds isolated from leaves of Diospyros kaki on stimulus-induced superoxide generation and tryrosyl phosphorylation of proteins in human neutrophils. Clinica Chimica Acta 326, 169–175. Chen, G., Xu, S.X., Sha, Y., 2000. Studies on the constituents of Diospyros kaki leaves. Chinese Journal of Medicinal Chemistry 10, 298–299. Chen, J., Zhao, F.L., Zhou, Q.J., 2010. Antimicrobial activities of volatile oil from persimmon leaves. Journal of Xiangfan University 31, 34–36. Chinese Pharmacopoeia Commission, 2012. The First Supplement of Chinese Pharmacopoeia. China Medical Science Press, Beijing. Chou, C.T., 1984. Studies on constituents of Diospyros kaki leaves (I). Kuo LI Chungkuo I Yao Yen Chiu So Yen Chiu Pao Kao 7, 117–140. Clark, C.J., Smith, G.S., 1990. Seasonal changes in the mineral nutrient content of persimmon leaves. Scientia Horticultural 42, 85–97. Deng, H., He, M., Li, J., Luo, X.Y., Huang, R.B., 2011. Hypoglycemic effect of persimmon leaf polysaccharide in diabetic mice induced by streptozotocin. Chinese Journal of Experimental Traditional Medical Formulae 17, 114–117. Deng, R.C., Zhang, W.S., Yang, H.J., 2004. Effect of leaf of persimmon oral solution on rats with acute myocardial ischemia. Chinese Journal of Information on TCM 11, 591–592. Fan, J.P., He, C.H., 2006. Simultaneous quantification of three major bioactive triterpene acids in the leaves of Diospyros kaki by high-performance liquid chromatography method. Journal of Pharmaceutical and Biomedical Analysis 41, 950–956. Fan, J.P., Zhang, R.F., Zhang, X.H., Zhu, J.H., Huang, J.Z., 2011. Separation of three triterpene acids in leaves of Diospyros kaki by high performance liquid chromatography using hydroxypropyl-β-cyclodextrin as mobile phase modifier. Journal of Liquid Chromatography & Related Technologies 34, 1340–1355. Funayama, S., Hikino, H., 1979. Hypertensive principles of Diospyros kaki leaves. Chemical & Pharmaceutical Bulletin 27, 2865–2868. Gibbons, E., 1962. Stalking the Wild Asparagus. Alan C Hood & Co., Chambersburg. Guo, M., Dong, X.P., 1999. Research summary of Diospyors kaki. Chongqing Chinese Traditional Herbal Drugs 2, 164. Han, J., Kang, S., Chou, S., Kim, H., Leem, K., Chung, S., Kim, C., Chung, J., 2002. Free radical scavenging effect of Diospyros kaki, Laminaria japonica and Undaria pinnatifida. Fitoterapia 73, 710–712. Han, K.H., Han, D.C., Zhang, Y.M., 1983. Pharmacological effect and clinical application of Diospyros kaki leaves. Research on Zhong Cheng Yao 7, 27–28. He, Y.L., Zhao, Y.L., Wang, X.Z., 1986. Determination of constituents in Hemostatic No. 4. China Journal of Chinese Materia Medica 11, 40–42. Hiromichi, O., 2002. Health Food & Nutrition. Oriental Medical Press, Tokyo. Hospital 58 of the Chinese People's Liberation ArmyThe experimental study and primary clinical effect observation of Diospyros kaki leaves. Chinese Traditional Herbal Drugs Communication 58, 8–10. Hu, J.F., Zuo, T.L., Zhang, H., Shen, Q., 2002. Characterization of the material content of Diospyros kaki leaves. Journal of Mongolia Polytechnic University 20, 61–64. Huang, S.L., Chen, L.F., Chen, D.Z., 1983. Alcohol extracts of Diospyros kaki leaves' influence on the heart function and hemodynamics of anaesthetic dogs. Guang Xi Yi Xue 5, 230–232. Ji, L.L., Zhang, Q.H., Cui, G.Y., 2003. Study on the antimicrobial activities of persimmon leaves against food spoilage and food-borne pathogens and related compounds. Food Science 24, 129–131. Ji, Z.P., Su, Y.Q., Lv, P.H., Zhang, C.L., 2006. Studies on chemical constituents and anti microbial activities of persimmon leaves. Chemistry and Industry of Forest Products 26, 87–90. Jung, U.J., Park, Y.B., Kim, S.R., Choi, M.S., 2012. Supplementation of persimmon leaf ameliorates hyperglycemia, dyslipidemia and hepatic fat accumulation in type 2 diabetic mice. PLoS One 7, e49030. Kawakami, K., Aketa, S., Sakai, H., Watanabe, Y., Nishida, H., Hirayama, M., 2011a. Antihypertensive and vasorelaxant effects of water-soluble proanthocyanidins from persimmon leaf tea in spontaneously hypertensive rats. Bioscience, Biotechnology, and Biochemistry 75, 1435–1439. Kawakami, K., Nishida, H., Tatewaki, N., Nakajima, Y., Konishi, T., Hirayama, M., 2011b. Persimmon leaf extract inhibits the ATM activity during DNA damage response induced by doxorubicin in A549 lung adenocarcinoma cells. Bioscience, Biotechnology, and Biochemistry 75, 650–655. Kawakami, K., Shibukura, Y., Kanno, T., Furuki, T., Aketa, S., Hirayama, M., 2011. Identification of 20 -galloylated flavonol 3-O-glycosides accumulating in developing leaves of persimmon. Phytochemical Analysis 22, 403–410. Kotani, M., Matsumoto, M., Fujita, A., Higa, S., Wang, W., Suemura, M., Kishimoto, T., Tanaka, T., 2000. Persimmon leaf extract and astragalin inhibit development of dermatitis and IgE elevation in NC/Nga mice. Journal of Allergy and Clinical Immunology 106, 159–166. Lan, M., 1975. Diannan Bencao. Yunnan People's Press, Yunnan. Lee, J.S., Lee, M.K., Ha, T.Y., Bok, S.H., Park, H.M., Jeong, K.S., Woo, M.N., Do, G.M., Yeo, J.Y., Choi, M.S., 2006. Supplementation of whole persimmon leaf improves lipid profiles and suppresses body weight gain in rats fed high-fat diet. Food and Chemical Toxicology 44, 1875–1883. Liang, C., Fu, F.M., 1985. Pharmacological effect on the cardiovascular system of Diospyros kaki leaves. Yao Xue Tong Bao 20, 245. Liang, W.B., Hu, C.S., She, X.W., 2000. Vc content and its distribution pattern in the leaves of 410 plants in hunan. Journal of central South Forestry University 20, 61–64.


67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132

12

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48


Lin, L., Chou, C., Chen, C., 1988. Constituents of Diospyros kaki leaves. Chung hua Yao Husea Tsa Chin 40, 195. Lin, S.R., Wang, J.L., Wu, X.N., 1996. Study on the antioxidant and antiatheroscloresis activity of Diospyrus kaki leaves tea. Strait Pharmaceutical Journal 8, 14–15. Liu, J., Lang, A.D., Nie, L., 2008. HPLC chromatographic fingerprints of persimmon leaves flavones. China Journal of Chinese Materia Medica 33, 575–576. Liu, L., Liu, R., Zhang, J., Zhang, Z., 2012. Study on the PEG-based microwave-assisted extraction of flavonoid compounds from persimmon leaves. Journal of Separation Science 35, 3412–3420. Mallavadhani, U.V., Panda, A.K., Rao, Y.R., 1998. Pharmacology and chemotaxonomy of Diospyros. Phytochemistry 49, 901–951. Matsumoto, M., Kotani, M., Fujita, A., Higa, S., Kishimoto, T., Suemura, M., Tanaka, T., 2002. Oral administration of persimmon leaf extract ameliorates skin symptoms and transepidermal water loss in atopic dermatitis model mice, NC/Nga. British Journal of Dermatology 146, 221–227. Matsura, S., Linuma, M., 1971. Components of the leaves of Diospyros kaki. Yakugaku Zasshi 90, 905. Mizuo, M., 1995. Japanese Herbal Medicines. The New Japanese Law press, Tokyo. Moon, S.H., 1996. Anti-tumor effect of persimmon leaves in vivo using sarcoma180. Journal of the Korean Society of Food Science and Nutrition 25, 865–870. Nakatani, M., Miyazaki, Y., Iwashitaa, T., Naokia, H., Hase, T., 1989. Triterpenes from Ilex rotunda fruits. Phytochemistry 28, 1479–1482. Ouyang, P., Bei, W.J., Lai, W.Y., Xu, D.L., Peng, W.L., 2003a. In vitro effects of flavone from leaves of Diospyros kaki on rat cardiac myocyte apoptosis in-duced by hypoxia-reoxygenation and advanced glycation end products. Journal of First Military Medicine University 23, 680–682. Ouyang, P., Bei, W.J., Lai, W.Y., Xu, D.P., Peng, W.L., 2003b. Effects of flavone from leaves of Diospyros kaki on adventitial fibroblast proliferation induced advanced glycation end-products in vitro. Journal of First Military Medicine University 23, 49–51. Ouyang, P., Zhang, B., Bei, W.J., Lai, W.Y., Peng, W.L., Xu, D.L., 2007. Effects of flavone from leaves of Diospyros kaki on expression of apoptosis signal-regulating kinase 1 and rat vascular smooth muscle cells proliferation by tumor necrosis factor-α in vitro. Journal of Chinese Medicinal Materials 30, 819–822. Qin, F.Z., Lin, X., Zhang, X.D., Huang, R.B., 2008. The hypotensive effects of persimmon leaf flavonoids. Modern Traditional Chinese Medicine 28, 61–63. Sa, Y.S., Kim, S.J., Choi, H.S., 2005. The anticoagulant fraction from the leaves of Diospyros kaki L. has an antithrombotic activity. Archives of Pharmacal Research 28, 667–674. Sakanaka, S., Tachibana, Y., Okada, Y., 2005. Preparation and antioxidant properties of extracts of Japanese persimmon leaf tea (kakinoha-cha). Food Chemistry 89, 569–575. Shi, J.Q., Huang, S.H., Shen, L.L., Zhang, J.M., 1999. The antioxidative activities of ethanolic extracts from persimmon leaves in lard. Science and Technology of Food Industry 20, 22–23. Sun, L., Zhang, J., Fang, K., Ding, Y., Zhang, L., Zhang, Y., 2014. Flavonoids from persimmon (Diospyros kaki) leaves (FPL) attenuate H2O2-induced apoptosis in MC3T3-E1 cells via the NF-[small kappa]B pathway. Food & Function 5, 471–479. Sun, L.J., Zhang, J.B., Lu, X.Y., Zhang, L.Y., Zhang, Y.L., 2011. Evaluation to the antioxidant activity of total flavonoids extract from persimmon (Diospyros kaki L.) leaves. Food and Chemical Toxicology 49, 2689–2696. Sun, Y., Tan, H.D., Lan, X.B., Huang, J.C., Huang, R.B., 2009. Protective effect of persimmon flavone pretreatment on rat myocardial ischemic and reperfusion Injury. Journal of Ji Ning Medical College 32, 9–11. Tezuka, M., Kuroyangi, M., Yoshihira, K., 1972. Naphthoquinone derivatives from the Ebenaceae IV. Naphthoquinone derivatives from Diosoyros kaki Thunb. and D. kaki Thunb var. sylvestris Makino. Chemical & Pharmaceutical Bulletin 20, 2029–2035. The National Compiling Group of Chinese Herbal MedicineCompiling of Chinese Herbal Medicine(II). People's Medical Publishing House, Beijing.

49 Thuong, P.T., Lee, C.H., Dao, T.T., Nguyen, P.H., Kim, W.G., Lee, S.J., Oh, W.K., 2008. Triterpenoids from the leaves of Diospyros kaki (persimmon) and their 50 inhibitory effects on protein tyrosine phosphatase 1B. Journal of Natural 51 Products 71, 1775–1778. 52 Tsurunaga, Y., Takabayashi, Y., Nishi, M., Suzuki, Y., 2011. Differences in the ascorbic acid, astragalin, and polyphenol contents, and the DPPH radical scavenging 53 activity of 22 commercial persimmon leaf tea products. Journal of Home 54 Economics of Japan 62, 437–444. 55 Wang, L., Xu, M.L., Rasmussen, S.K., Wang, M.H., 2011. Vomifoliol 9-O-α56 arabinofuranosyl (1-6)-β-d-glucopyranoside from the leaves of Diospyros kaki stimulates the glucose uptake in HepG2 and 3T3-L1 cells. Carbohydrate 57 Research 346, 1212–1216. 58 Wang, S.S., Zong, W.P., Xu, D., 2004. Effect of persimmon leaves on metrorrhagia by 59 drug abortion of pregnant rats on early and middle stages. Journal of Hebei 60 Traditional Chinese Medicine and Pharmacology 19, 1–4. Wu, R., Qin, R.A., Yin, R.J., Wang, D.Q., Li, C.Y., 2012. Experimental study on acute 61 toxicity and genetic toxicity of Diospyros kaki extract. World Science and 62 Technology/Modernization of Traditional Chinese Medicine and Materia Med63 ica 14, 2201–2204. 64 Wu, X.N., Wang, J.L., 1999. Study on the actions of banting and reducing blood lipid of fresh leaf of persimmon's juice with experimental hyperlipidemia rats. China 65 Public Health 15, 302–303. 66 Xiong, W.Y., Wang, J.Z., Shi, T.D., 1993. Woody Medicine Plants of China. Shanghai 67 Education & Technology Press, Shanghai. Xue, Y.L., Miyakawa, T., Hayashi, Y., Okamoto, K., Hu, F., Mitani, N., Furihata, K., 68 Sawano, Y., Tanokura, M., 2011. Isolation and tyrosinase inhibitory effects of 69 polyphenols from the leaves of persimmon Diospyros kaki. Journal of Agricul70 tural and Food Chemistry 59, 6011–6017. 71 Yang, J.X., Yuan, J.F., Li, F.R., 2003. The anti-oxidative effects of persimmon leaf flavonoid in vitro. Acta Nutrimenta Sinica 25, 215–217. 72 Ye, G., 1933. Bencao Zaixin. Shanghai Qunxue Publishing House, Shanghai. 73 Yu, F.H., Feng, J., Wang, S.S., Yan, P., 2005. Influence of Diospyros kaki leaf on NO, 74 NOS and ET in uterus homogenate of rats after medicine induced abortion. 75 Modern Journal of Integrated Traditional Chinese and Western Medicine 14, 3205–3206. 76 Yu, Y.Z., Yu, Z.Y., Guo, J., 1988. The experimental study and primary clinical effect 77 observation of Diospyros kaki leaves the treatment of ischemic cerebrovascular 78 disease. Medical Journal of Chinese People's Liberation Army 13, 30–31. Yuan, J., Yang, J., Zhang, Z., 2006. Determination of rutin and oleanolic acid in the 79 leaves of persimmon in different seasons by RP-HPLC (2013-1026). Natural Q11 80 Product Research and Development 18. 81 Zakaria, M., Jeffieys, J., Materman, P., 1984. Nahthoquinones and triterpenes from 82 some Asian Diospyros species. Phytoehemistry 23, 1481–1484. Zeng, X.Y., Chen, X.F., Li, Y.D., 1987. Effect of rats' immunity from persimmon leaves 83 extract- DK-N3. Guangxi Medical Journal 9, 125–127. 84 Zhang, J.X., Zhou, J.K., Wu, Z.Y., 2002. Determination of organic acids and inorganic 85 anions in persimmon leaves by single-column ion chromatography. Physical 86 Testing and Chemical Analysis 38, 29–31. Zhang, S.H., Wang, Y.Z., Meng, F.Y., Li, Y.L., Li, C.X., Duan, F.P., Wang, Q., Zhang, X.T., 87 Zhang, C.N., 2014. Studies of the microbial metabolism of flavonoids extracted Q12 88 from the leaves of Diospyros kaki by intestinal bacteria. Archives of Pharmacal 89 Research , http://dx.doi.org/10.1007/s12272-014-0421-6. Zhou, F.X., Liang, P.Y., Wen, J., Ma, Y., 1987. Studies on the costituents of Diospyros 90 kaki leaves. China Journal of Chinese Materia Medica 12, 38. 91 Zou, S.Q., Sun, W., 2009. Simultaneous determination of oleanolic acid and ursolic 92 acid in persimmon leaves by RP-HPLC. Chinese Journal of Analysis Laboratory 93 28, 18–21. Zu, S., Lei, Y.Y., 1989. The feeding value of Diospyros kaki leaves' powder. Feed 94 Research 11, 27–29. 95


Clerodendrum serratum (L.) Moon. - a review on traditional uses, phytochemistry and pharmacological activities.

Traditional uses, phytochemistry and pharmacological properties of the genus Peucedanum: a review.

Persimmon (Diospyros kaki) fruit: hidden phytochemicals and health claims.

Persimmon Leaves (Diospyros kaki) Extract Protects Optic Nerve Crush-Induced Retinal Degeneration.

Extraction, Identification and Photo-Physical Characterization of Persimmon (Diospyros kaki L.) Carotenoids.

Production of dodecaploid plants of Japanese persimmon (Diospyros kaki L.) by colchicine treatment of protoplasts.

Traditional uses, phytochemistry and pharmacology of Ficus carica: a review.

Ficus carica L. (Moraceae): Phytochemistry, Traditional Uses and Biological Activities.

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

Mineral profile of kaki fruits (Diospyros kaki L.).

The genus Psiadia: Review of traditional uses, phytochemistry and pharmacology.

Sulphated modification of a polysaccharide obtained from fresh persimmon (Diospyros kaki L.) fruit and antioxidant activities of the sulphated derivatives.

Portulaca oleracea L.: a review of phytochemistry and pharmacological effects.

The genus spilanthes ethnopharmacology, phytochemistry, and pharmacological properties: a review.

Muntingia calabura: a review of its traditional uses, chemical properties, and pharmacological observations.

Jatropha gossypiifolia L. (Euphorbiaceae): A Review of Traditional Uses, Phytochemistry, Pharmacology, and Toxicology of This Medicinal Plant.

Citrullus colocynthis (L.) Schrad (bitter apple fruit): a review of its phytochemistry, pharmacology, traditional uses and nutritional potential.

Review on medicinal uses, pharmacological, phytochemistry and immunomodulatory activity of plants.

Accelerated solvent extraction of carotenoids from: Tunisian Kaki (Diospyros kaki L.), peach (Prunus persica L.) and apricot (Prunus armeniaca L.).

Alepidea amatymbica Eckl. & Zeyh.: A Review of Its Traditional Uses, Phytochemistry, Pharmacology, and Toxicology.

Traditional uses, phytochemistry, pharmacology, toxicology and quality control of Alisma orientale (Sam.) Juzep: a review.

Radix Bupleuri: A Review of Traditional Uses, Botany, Phytochemistry, Pharmacology, and Toxicology.

Traditional Uses, Phytochemistry, and Pharmacology of Olea europaea (Olive).

Effect of Persimmon Peel (Diospyros kaki Thumb.) Extracts on Lipid and Protein Oxidation of Raw Ground Pork During Refrigerated Storage.