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Pdf 161.55 K Iranian Journal of Pharmaceutical Research (2011), 10 (2): 261-264 Copyright © 2011 by School of Pharmacy Received: July 2009 Shaheed Beheshti University of Medical Sciences and Health Services Accepted: July 2009 Original Article Screening of Korean Medicinal Plant Extracts for Į-Glucosidase Inhibitory Activities Shruti Sanchetia, Sandesh Sanchetia 6HXQJ+XQ/HHa, -DH(XQ/HHa and Sung-Yum Seoa,b* aDepartment of Biology, Kongju National University, Kongju 314-701, Korea. bKorean Collection of Herbal Extract, Inc., Kongju 314-701, Korea. Abstract Glycosidases are the enzymes involved in various biochemical processes related to metabolic disorders and diseases. Therefore, much effort has been focused on searching novel pharmacotherapy for the treatment of these ailments from medicinal plants due to higher safety margins. To pursue these efforts, the present study was performed to evaluate the Į-glucosidase inhibitory activities of thirty Korean medicinal plant extracts. Among the plants studied, Euonymus sachalinensis, Rhododendron schlippenbachii, Astilbe chinensis and Juglans regia showed the strongest Į-glucosidase inhibitory activity with IC50YDOXHVRIDQGJ P/UHVSHFWLYHO\,QDGGLWLRQMeliosma oldhamii and Symplocos chinensis showed moderate Į-glucosidase inhibition with IC50YDOXHVRIDQGJP/UHVSHFWLYHO\7KHUHIRUHWKH\ might prove to be a potential natural source for the treatment of metabolic ailments such as, diabetes, and need further investigations. Keywords: Į-Glucosidase inhibitor; Korean plants; Screening; Glycosidases. Introduction diabetes, obesity, glycosphingolipid lysosomal storage disease, HIV infections, and tumors Glycosidases are widespread in (1-3). These observations indicate that the microorganisms, plants, and animals. They inhibition of glycosidases would represent a are a very important class of enzymes, which novel pharmacological approach towards the catalyze a hydrolytic cleavage of glycosidic treatment of the above mentioned complications, bonds in oligosaccharides or glycoconjugates. including diabetes. Among these glycosidases, Į-glucosidase is Diabetes mellitus (DM) is a metabolic able to catalyze the cleavage of glycosidic bonds disorder characterized by chronic hyperglycemia involving terminal glucose connected at the site WKDW KDV D VLJQL¿FDQW LPSDFW RQ WKH KHDOWK of cleavage through Į-linkage at the anomeric quality of life and life expectancy of patients, center (1-3). as well as health care system (1, 4). In clinical Glycosidases are involved in several practice, the DM treatment is restricted to the important biological processes (like: digestion, use of oral hypoglycemic agents and insulin, biosynthesis of glycoproteins and lysosomal where the former possesses serious side effects catabolism of glucoconjugates) related to (5). Therefore, many traditional herbal remedies metabolic disorders and diseases, such as, for treating diabetes used throughout the world as plant drugs and herbal formulations, are * Corresponding author: frequently considered to be free from side effects E-mail: [email protected] and less toxic than the synthetic one (6). Thus, Sancheti Sh et al. / IJPR (2011), 10 (2): 261-264 Table 1. Į-Glucosidase inhibitory activities and IC50 values of the studied Korean plant extracts. Į-glucosidase inhibition Plant name Family Plant part a % Inhibition IC50 Values (μg/mL) 1 Actinidia arguta Actinidiaceae Whole plant 33 ± 1 -- 2 Meliosma oldhamii Sabiaceae Whole plant 67 ± 3 150 ± 3 3 Aster tataricus Asteraceae Stem n. a. -- 4 Capsella bursa-pastoris Brassicaceae Whole plant 23 ± 1 -- 5 Allium macrostemon Alliaceae Whole plant 19 ± 2 -- 6 Rhododendron schlippenbachii Ericaceae /HDI 98 ±1 20 ± 1 7 Pyrola japonica Pyrolaceae Whole plant 29 ± 2 -- 8 Symplocos chinensis Symplocaceae /HDI 52 ± 2 220 ± 5 9 Juglans regia Juglandaceae Whole plant 99 ± 1 80 ± 2 10 Sinapsis alba Brassicaceae Seed 22 ± 2 -- 11 Aster tataricus Asteraceae Root 17 ± 3 -- 12 Magnolia kobus Magnoliaceae Whole plant 4 ± 1 -- 13 Inula helenium Asteraceae Whole plant 9 ± 2 -- 14 Digitaria violascens Poaceae Whole plant n. a. -- 15 Dendropanax morbifera Araliaceae Whole plant 39 ± 3 -- 16 Sesamum indicum Pedaliaceae Whole plant 11 ± 2 -- 17 Alisma canaliculatum Alismataceae Rhizomes 22 ± 3 -- 18 Celtis sinensis Cannabaceae Whole plant 25 ± 1 -- 19 Astilbe chinensis Saxifragaceae Rhizomes 95 ± 2 30 ± 2 20 Corydalis remota Papaveraceae Whole plant 23 ± 2 -- 21 Phlomis umbrosa /DELDWDH Whole plant 12 ± 2 -- 22 Curcuma zedoaria Zingiberaceae Whole plant 27 ± 2 -- 23 Gleditsia japonica Fabaceae Whole plant n. a. -- 24 Miscanthus sinensis Poaceae Whole plant 30 ± 3 -- 25 +HUDFOHXPPRHOOHQGRUI¿L Apiaceae Whole plant 17 ± 2 -- 26 Draba nemorosa Brassicaceae Whole plant 11 ± 1 -- 27 Vaccinium hirtum Ericaceae Whole plant 46 ± 1 -- 28 Smilax sieboldii Smilacaceae Whole plant 24 ± 3 -- 29 Euonymus sachalinensis Celastraceae /HDI 99 ± 1 10 ± 1 30 Sinomenium acutum Menispermaceae Bark 28 ± 2 -- Note: The IC50 values of 1,2,3,4,6-penta-O-galloyl--ȕ-D-glucose (positive control) for Į-glucosidase inhibitory activity was measured DVJP/UHVSHFWLYHO\ a All results are represented as mean ± SD (n = 3). n. a.: no activity. --: Not determined (IC50 YDOXHVZHUHRQO\GHWHUPLQHGIRUWKHSODQWH[WUDFWVZLWKLQKLELWLRQDWPJP/ many plants and crude drugs have recently screened thirty Korean medicinal plants. The been tested for their effects on Į-glucosidase GHWDLOVRIWKHSODQWV¶VFLHQWL¿FQDPHVDQGIDPLOLHV inhibition. are listed in Table 1. A literature survey did not 7R SXUVXH WKH ¿QGLQJV LQ WKLV UHVHDUFK ZH show any reference to a previous work on the 262 Screening of Korean Medicinal Plant Extracts Į-glucosidase inhibitory activities of these thirty by the amount of p-nitrophenol released in the plant extracts. reaction mixture at 410 nm using microplate reader. Individual blanks were prepared for Experimental correcting the background absorbance, where the enzymes were replaced with buffer. Controls Plant material were conducted in an identical manner replacing The dried and matured plant parts of thirty the plant extracts with methanol. 1, 2, 3, 4, Korean medicinal herbs were obtained from 6-penta-O-galloyl--ȕ-D-glucose was used as ³.RUHDQ &ROOHFWLRQ RI +HUEDO ([WUDFWV´ D positive control. All experiments were carried Biotech company in Korea. A collection out in triplicates. The inhibition percentage of of voucher specimen is available with Į-glucosidase was assessed by the following the company (Korea Collection of Herbal formula: Extracts, 2000). I ĮJOXFRVLGDVH ; ǻ$Control ǻ$Sample) Extraction ǻ$Control The dried plant parts individually were chopped into small pieces and pulverized into a ǻ$Control ǻ$Testt ǻ$Blank ¿QHSRZGHU7KHSRZGHUHGSODQWPDWHULDOV g, dry weight) were kept for extensive decoction ǻ$Sample ǻ$Testtǻ$Blank in 80% methanol for 3 days at room temperature. The extracts were then concentrated using rotary Statistical analyses vacuum evaporator at 20-30& to obtain the dried All assays were performed at least three times crude extracts. with triplicate samples. All results are expressed as mean ± SD. IC50 values were only determined Reagents IRU WKH SODQW H[WUDFWV ZLWK LQKLELWLRQ DW Į-Glucosidase (from Saccharomyces PJP/E\SORWWLQJDSHUFHQWLQKLELWLRQYHUVXV cerevisiae type I) and 4-nitrophenyl Į-D- concentration curve, in which the concentration glucopyranoside were purchased from of sample required for 50% inhibition was 6LJPD$OGULFK 6W /RXLV 02 86$ 2WKHU determined and expressed as IC50 value. commercially available reagents and solvents were used as received. Results and Discussion Į*OXFRVLGDVHDVVD\ Plants have always been used as an exemplary The enzyme inhibition activity for source of drugs and many of the currently Į-glucosidase was evaluated according to the available drugs have been directly or indirectly method previously reported by Shibano et al. derived from them (8). Many herbal extracts ZLWKPLQRU PRGL¿FDWLRQV 7KH UHDFWLRQ are being used in the preparation of advanced PL[WXUHFRQVLVWHGRIȝ/RI0SKRVSKDWH remedies for diabetes, in which Į-glucosidase EXIIHU ZLWK S+ RI ȝ/ RI P0 inhibitors play an important role by controlling 4-nitrophenyl Į-D-glucopyranoside (dissolved postprandial blood glucose levels by means of in 0.1 M phosphate buffer, with pH of 7.0), 10 retarding uptake of dietary carbohydrates (9). ȝ/ RI WHVW VDPSOH DQG ȝ/ RI Į-glucosidase Therefore, in search of such potent Į-glucosidase VROXWLRQ D VWRFN VROXWLRQ RI PJP/ LQ inhibitors from natural source, in the present M phosphate buffer, with pH of 7.0 was diluted study thirty Korean medicinal plant extracts have WR 8QLWP/ ZLWK WKH VDPH EXIIHU ZLWK S+ been evaluated for their Į-glucosidase inhibitory of 7.0 just before assay). This reaction mixture activity and compared with 1, 2, 3, 4, 6-penta-O- ZDV WKHQ LQFXEDWHG DW & IRU PLQ7KHQ galloyl--ȕ-D-glucose as a positive control. the reaction was terminated by the addition of In this study, the Į-glucosidase inhibitory ȝ/RI0VRGLXPFDUERQDWHVROXWLRQ7KH activity of the thirty plant extracts was evaluated enzymatic hydrolysis of substrate was monitored DW PJP/ FRQFHQWUDWLRQ DW WKH SUHOLPLQDU\ 263 Sancheti Sh et al. / IJPR (2011), 10 (2): 261-264 Table 2. Details of the traditional indications of the active plant extracts. Sr. no. Plant name Traditional indications 1 Meliosma oldhamii Treatment of ailments of the liver 2 Rhododendron schlippenbachii Discharge of cardiotonic phlegm 3 Symplocos chinensis &ROGIHYHUPDODULDUHOLHIRIFRXJKGHWR[L¿FDWLRQ 4 Juglans regia $QWLLQÀDPPDWRU\DVWULQJHQWDQWLFDQFHUEORRGSXUL¿HUOD[DWLYHGLXUHWLFDQWKHOPLQWLF
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