Letters to the Editor
In vitro study on α‑amylase inhibitory activity of an Ayurvedic medicinal plant, Anacyclus pyrethrum DC root Access this article online Website: www.ijp‑online.com
Quick Response Code:
DOI: 10.4103/0253-7613.132204
Table 1: α‑amylase inhibitory effects of different extracts of Anacyclus pyrethrum DC root Plant extracts
Concentration (µg/ml)
Percentage of inhibition
IC50 value (µg/ml)
Acarbose
10 20 40 60 80 100 10 20 40 60 80 100 10 20 40 60 80 100 10 20 40 60 80 100 10 20 40 60 80 100 10 20 40 60 80 100
14.19 19.75 27.15 34.56 46.90 51.23 24.68 39.50 56.16 64.19 70.36 78.38 19.75 29.00 47.52 60.49 66.04 70.36 17.89 29.00 43.82 56.16 63.57 68.51 29.00 40.11 66.04 78.38 83.33 88.26 20.98 30.86 43.82 58.01 70.98 82.09
92.38±3.229
Sir, Diabetes mellitus (DM) consists of a group of metabolic diseases characterized by inappropriate hyperglycemia resulting from defects in insulin secretion, insulin action or both. Perhaps food habits and genetic factors play a major role for diabetes. A study revealed that urbanization of rural India has doubled the rate of diabetes, in which type 2 DM is the primary threat to human health due to the increasing prevalence, chronic course and disabling complications.[1] The medicinal plants or natural products involve retarding the absorption of glucose by inhibiting the carbohydrate hydrolyzing enzymes, such as pancreatic amylase. The inhibition of this enzyme delays carbohydrate digestion and prolong overall carbohydrate digestion time, resulting in the reduction in glucose absorption rate and consequently dulling the postprandial plasma glucose rise. Several indigenous medicinal plants have high potential in inhibiting α‑amylase enzyme activity.[2] Different parts of plant have been used predominantly in folk medicine worldwide for the treatment of various diseases such as paralysis, epilepsy, fever, pharyngitis, tonsillitis, and diabetes.[3] However, their antidiabetic activity has not been documented. This study was carried out to evaluate in vitro inhibitory effects of various extracts (petroleum ether, chloroform, ethyl acetate, acetone, ethanol, and water) of A. pyrethrum root on porcine pancreatic amylase activity. The species for the proposed study, A. pyrethrum DC root was purchased from local market (M.A.S. Stores, Country drugs wholesale and retail, Erode, India) and authenticated by Prof. P. Jayaraman, Director, National Institute of Herbal Science, Chennai‑45, (Ref. no: PARC/2011/896). The dried powdered plant root of A. pyrethrum DC was extracted using a soxhlet apparatus sequentially with petroleum ether, chloroform, ethyl acetate, acetone, ethanol, and water. Each extract was evaporated using rotary evaporator under reduced pressure. The preliminary phytochemical screening of extracts was carried out by chemical tests.[4] The dried extracts were dissolved in dimethyl sulfoxide to make different concentrations and subjected to α‑amylase inhibitory assay[5] and the results were tabulated [Table 1]. 350 Indian Journal of Pharmacology | June 2014 | Vol 46 | Issue 3
Chloroform
Ethyl acetate
Acetone
Ethanol
Water
40.34±4.173
52.52±4.443
57.29±3.996
29.25±2.897
49.36±1.010
In comparison of IC 50 value of α‑amylase inhibitory effect of different extracts of A. pyrethrum DC root against porcine pancreatic amylase revealed that ethanol extract at higher concentration showed 88.26% (IC50‑29.25 µg/ml) significant α‑amylase inhibitory effect than the other extracts. Simultaneously, all extract showed appreciable α‑amylase
Letters to the Editor
inhibitory activity except petroleum ether extract when compared with acarbose. It may be due to the presence of chemical constituents such as alkaloids, flavonoids, phytosteroids, and also glycosides. The plant‑based α‑amylase inhibitory offers a prospective therapeutic approach for the management of diabetes. [6] This study supports the Ayurvedic concept that A. pyrethrum DC root could be useful in management of diabetes.[7] Furthermore, the in vivo antidiabetic activity of these extracts needs to be assessed prior to clinical use. V. Kishor Kumar, K. G. Lalitha1 Department of Phytopharmacy and Phytomedicine, J. K. K. Munirajah Medical Research Foundation’s, Annai J. K. K Sampoorani Ammal College of Pharmacy, B. Komarapalayam, Namakkal, 1Department of Pharmaceutical Chemistry, Ultra College of Pharmacy, Madurai, Tamil Nadu, India
Correspondence to: Dr. K. G. Lalitha, E‑mail: [email protected]
References 1. Mitra A. Effect of a composite of tulsi leaves, amla, bitter gourd, gurmur leaves, jamun fruit and seed in type 2 diabetic patients. J Clin Diagn Res 2007;6:511‑20. 2. Prashanth D, Padmaja R, Samiulla DS. Effect of certain plant extracts on alpha‑amylase activity. Fitoterapia 2001;72:179‑81. 3. Anonymous. The Wealth of India: Raw Materials. Vol. I. New Delhi: CSIR Press; 1985. p. 248. 4. Khandelwal KR. Practical Pharmacognosy Techniques and Experiments. Pune, India: Nirali Prakashan; 2000. p. 149‑54. 5. Tamil IG, Dineshkumar B, Nandhakumar M, Senthilkumar M, Mitra A. In vitro study on α‑amylase inhibitory activity of an Indian medicinal plant, Phyllanthus amarus. Indian J Pharmacol 2010;42:280‑2. 6. McCue P, Vattem D, Shetty K. Inhibitory effect of clonal oregano extracts against porcine pancreatic amylase in vitro. Asia Pac J Clin Nutr 2004;13:401‑8. 7. Anonymous. The Ayurvedic Pharmacopoeia of India‑Part I. 1st ed. New Delhi: The Controller of Publications; 1999. p. 1‑2.
Noninvasive measurement of systolic blood pressure in rats: A novel technique Access this article online Website: www.ijp‑online.com
Quick Response Code:
DOI: 10.4103/0253-7613.132207
Sir, Essential hypertension affects 95% of hypertensive patients worldwide countries.[1] Animal models are required to understand the physiology of essential hypertension. Intraarterial cannulation is generally considered the most physiological method of blood pressure (BP) recording in animals like rats. The procedure of arterial cannulation in small animals and to maintain the patency of arterial catheter for long time experimentation is very difficult and time consuming.[2,3] Various studies has reported the strong correlation between tail‑cuff and intraarterial BPs measured simultaneously in conscious rats.[4,5] These comparisons have provided important validations of the BP recorded by tail‑cuff method. However, there are various factors including heating and restraint that can alter the BP recorded by noninvasive methods.[6] Photoplethysmography (PPG), piezoplethysmography and volume pressure recording used in various noninvasive blood pressure (NIBP) techniques for measuring the BP in small animals like a rat and mice are expensive. Systolic BP can be determined by measuring the pressure value in the cuff when PPG pulse reappears during deflation. This is a simple technique and does not need calculations or formulae.[7] In view of the above, we hypothesized that BP in rat can be measured by using pulse
transducer with physiograph and an appropriate rat tail‑cuff. In this study, we recorded the systolic BP of rats by an innovated device reproducible and that correlates strongly with BP measured subsequently by NIBP machine from ADinstruments (ADI) (IN125NIBP controller) Australia. This experimental design has the advantage of being simple, convenient and of low cost. The study was conducted on albino rats (n = 6) weighing 180-250 g obtained from central animal house of King George’s Medical University, Lucknow. BP was measured by two devices at 9:30 am daily for 5 days. The device was developed at the Department of Physiology KGMU, Lucknow. Study was done in accordance with the CPCSEA guidelines. Sphygmomanometer, physiograph with coupler, pulse transducer, triway and noncollapsible rubber tubes were procured from various laboratories and inflatable rat tail‑cuff was designed by the authors [Figure 1]. The cuff consists of latex balloon measuring 5 cm × 2 cm with 0.5 mm thickness. This balloon was placed in a circular plastic case having a diameter of 23 mm with a central hole of 12 mm diameter. The balloon was kept in such a fashion that it remains in contact with an inner surface of plastic case around the central hole, so that this balloon encircles the tail. One end of the triway was connected with the balloon (tail‑cuff) and other two ends were connected to inflating‑deflating pump and sphygmomanometer [Figure 2]. The system measures systolic BP by determining the cuff pressure (reflected on sphygmomanometer) at which blood flow (pulse) to the tail was eliminated. This elimination of blood flow (pulse) was recorded by pulse transducer connected to single‑channel physiograph through a suitable coupler. The animals were kept in restrainers after acclimatizing them, the tail was passed through the hole of the newly designed cuff and the pulse transducer was tied around the tail distal to Indian Journal of Pharmacology | June 2014 | Vol 46 | Issue 3 351
Copyright of Indian Journal of Pharmacology is the property of Medknow Publications & Media Pvt. Ltd. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use.
In vitro study on α‑amylase inhibitory activity of an Ayurvedic medicinal plant, Anacyclus pyrethrum DC root Access this article online Website: www.ijp‑online.com
Quick Response Code:
DOI: 10.4103/0253-7613.132204
Table 1: α‑amylase inhibitory effects of different extracts of Anacyclus pyrethrum DC root Plant extracts
Concentration (µg/ml)
Percentage of inhibition
IC50 value (µg/ml)
Acarbose
10 20 40 60 80 100 10 20 40 60 80 100 10 20 40 60 80 100 10 20 40 60 80 100 10 20 40 60 80 100 10 20 40 60 80 100
14.19 19.75 27.15 34.56 46.90 51.23 24.68 39.50 56.16 64.19 70.36 78.38 19.75 29.00 47.52 60.49 66.04 70.36 17.89 29.00 43.82 56.16 63.57 68.51 29.00 40.11 66.04 78.38 83.33 88.26 20.98 30.86 43.82 58.01 70.98 82.09
92.38±3.229
Sir, Diabetes mellitus (DM) consists of a group of metabolic diseases characterized by inappropriate hyperglycemia resulting from defects in insulin secretion, insulin action or both. Perhaps food habits and genetic factors play a major role for diabetes. A study revealed that urbanization of rural India has doubled the rate of diabetes, in which type 2 DM is the primary threat to human health due to the increasing prevalence, chronic course and disabling complications.[1] The medicinal plants or natural products involve retarding the absorption of glucose by inhibiting the carbohydrate hydrolyzing enzymes, such as pancreatic amylase. The inhibition of this enzyme delays carbohydrate digestion and prolong overall carbohydrate digestion time, resulting in the reduction in glucose absorption rate and consequently dulling the postprandial plasma glucose rise. Several indigenous medicinal plants have high potential in inhibiting α‑amylase enzyme activity.[2] Different parts of plant have been used predominantly in folk medicine worldwide for the treatment of various diseases such as paralysis, epilepsy, fever, pharyngitis, tonsillitis, and diabetes.[3] However, their antidiabetic activity has not been documented. This study was carried out to evaluate in vitro inhibitory effects of various extracts (petroleum ether, chloroform, ethyl acetate, acetone, ethanol, and water) of A. pyrethrum root on porcine pancreatic amylase activity. The species for the proposed study, A. pyrethrum DC root was purchased from local market (M.A.S. Stores, Country drugs wholesale and retail, Erode, India) and authenticated by Prof. P. Jayaraman, Director, National Institute of Herbal Science, Chennai‑45, (Ref. no: PARC/2011/896). The dried powdered plant root of A. pyrethrum DC was extracted using a soxhlet apparatus sequentially with petroleum ether, chloroform, ethyl acetate, acetone, ethanol, and water. Each extract was evaporated using rotary evaporator under reduced pressure. The preliminary phytochemical screening of extracts was carried out by chemical tests.[4] The dried extracts were dissolved in dimethyl sulfoxide to make different concentrations and subjected to α‑amylase inhibitory assay[5] and the results were tabulated [Table 1]. 350 Indian Journal of Pharmacology | June 2014 | Vol 46 | Issue 3
Chloroform
Ethyl acetate
Acetone
Ethanol
Water
40.34±4.173
52.52±4.443
57.29±3.996
29.25±2.897
49.36±1.010
In comparison of IC 50 value of α‑amylase inhibitory effect of different extracts of A. pyrethrum DC root against porcine pancreatic amylase revealed that ethanol extract at higher concentration showed 88.26% (IC50‑29.25 µg/ml) significant α‑amylase inhibitory effect than the other extracts. Simultaneously, all extract showed appreciable α‑amylase
Letters to the Editor
inhibitory activity except petroleum ether extract when compared with acarbose. It may be due to the presence of chemical constituents such as alkaloids, flavonoids, phytosteroids, and also glycosides. The plant‑based α‑amylase inhibitory offers a prospective therapeutic approach for the management of diabetes. [6] This study supports the Ayurvedic concept that A. pyrethrum DC root could be useful in management of diabetes.[7] Furthermore, the in vivo antidiabetic activity of these extracts needs to be assessed prior to clinical use. V. Kishor Kumar, K. G. Lalitha1 Department of Phytopharmacy and Phytomedicine, J. K. K. Munirajah Medical Research Foundation’s, Annai J. K. K Sampoorani Ammal College of Pharmacy, B. Komarapalayam, Namakkal, 1Department of Pharmaceutical Chemistry, Ultra College of Pharmacy, Madurai, Tamil Nadu, India
Correspondence to: Dr. K. G. Lalitha, E‑mail: [email protected]
References 1. Mitra A. Effect of a composite of tulsi leaves, amla, bitter gourd, gurmur leaves, jamun fruit and seed in type 2 diabetic patients. J Clin Diagn Res 2007;6:511‑20. 2. Prashanth D, Padmaja R, Samiulla DS. Effect of certain plant extracts on alpha‑amylase activity. Fitoterapia 2001;72:179‑81. 3. Anonymous. The Wealth of India: Raw Materials. Vol. I. New Delhi: CSIR Press; 1985. p. 248. 4. Khandelwal KR. Practical Pharmacognosy Techniques and Experiments. Pune, India: Nirali Prakashan; 2000. p. 149‑54. 5. Tamil IG, Dineshkumar B, Nandhakumar M, Senthilkumar M, Mitra A. In vitro study on α‑amylase inhibitory activity of an Indian medicinal plant, Phyllanthus amarus. Indian J Pharmacol 2010;42:280‑2. 6. McCue P, Vattem D, Shetty K. Inhibitory effect of clonal oregano extracts against porcine pancreatic amylase in vitro. Asia Pac J Clin Nutr 2004;13:401‑8. 7. Anonymous. The Ayurvedic Pharmacopoeia of India‑Part I. 1st ed. New Delhi: The Controller of Publications; 1999. p. 1‑2.
Noninvasive measurement of systolic blood pressure in rats: A novel technique Access this article online Website: www.ijp‑online.com
Quick Response Code:
DOI: 10.4103/0253-7613.132207
Sir, Essential hypertension affects 95% of hypertensive patients worldwide countries.[1] Animal models are required to understand the physiology of essential hypertension. Intraarterial cannulation is generally considered the most physiological method of blood pressure (BP) recording in animals like rats. The procedure of arterial cannulation in small animals and to maintain the patency of arterial catheter for long time experimentation is very difficult and time consuming.[2,3] Various studies has reported the strong correlation between tail‑cuff and intraarterial BPs measured simultaneously in conscious rats.[4,5] These comparisons have provided important validations of the BP recorded by tail‑cuff method. However, there are various factors including heating and restraint that can alter the BP recorded by noninvasive methods.[6] Photoplethysmography (PPG), piezoplethysmography and volume pressure recording used in various noninvasive blood pressure (NIBP) techniques for measuring the BP in small animals like a rat and mice are expensive. Systolic BP can be determined by measuring the pressure value in the cuff when PPG pulse reappears during deflation. This is a simple technique and does not need calculations or formulae.[7] In view of the above, we hypothesized that BP in rat can be measured by using pulse
transducer with physiograph and an appropriate rat tail‑cuff. In this study, we recorded the systolic BP of rats by an innovated device reproducible and that correlates strongly with BP measured subsequently by NIBP machine from ADinstruments (ADI) (IN125NIBP controller) Australia. This experimental design has the advantage of being simple, convenient and of low cost. The study was conducted on albino rats (n = 6) weighing 180-250 g obtained from central animal house of King George’s Medical University, Lucknow. BP was measured by two devices at 9:30 am daily for 5 days. The device was developed at the Department of Physiology KGMU, Lucknow. Study was done in accordance with the CPCSEA guidelines. Sphygmomanometer, physiograph with coupler, pulse transducer, triway and noncollapsible rubber tubes were procured from various laboratories and inflatable rat tail‑cuff was designed by the authors [Figure 1]. The cuff consists of latex balloon measuring 5 cm × 2 cm with 0.5 mm thickness. This balloon was placed in a circular plastic case having a diameter of 23 mm with a central hole of 12 mm diameter. The balloon was kept in such a fashion that it remains in contact with an inner surface of plastic case around the central hole, so that this balloon encircles the tail. One end of the triway was connected with the balloon (tail‑cuff) and other two ends were connected to inflating‑deflating pump and sphygmomanometer [Figure 2]. The system measures systolic BP by determining the cuff pressure (reflected on sphygmomanometer) at which blood flow (pulse) to the tail was eliminated. This elimination of blood flow (pulse) was recorded by pulse transducer connected to single‑channel physiograph through a suitable coupler. The animals were kept in restrainers after acclimatizing them, the tail was passed through the hole of the newly designed cuff and the pulse transducer was tied around the tail distal to Indian Journal of Pharmacology | June 2014 | Vol 46 | Issue 3 351
Copyright of Indian Journal of Pharmacology is the property of Medknow Publications & Media Pvt. Ltd. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use.
