2.1 Chemicals Glucose, Glucose oxidase, Potassium mono hydrogen phosphate, Hydrochloric acid, Potassium dihydrogen phosphate, and EDTA were purchased from Merck and S igma. All chemicals and reagents used were of the analyti cal grade and highest commercially available purity. 2.2 Plant collection Leaves of Dendrocalamus hamiltonii (DH) , Dendrocalamus sikkimensis (DS), Bambusa balcooa (BB) , Bambusa pallida (BP) , Bambusa vulgaris (BV) were collected from the Sukna forest after taxonomic identification and authentic ation. The live accessions are deposited at the Bambusetum in K urseong Research Range, Sukna, Siliguri 2.3 Extract Preparation After shade dried (Temp<40?C.), plant material was g rounded into a moderately coarse powder was boiled with six teen parts of methanol for a period of 15 minutes 10 and filtered was evaporated under reduced pressure and dried .The yi eld of the extract was 1.2-2.06% (w/w). 2.4 Procedure for glucose oxidase method Trinder (1969) 12 method with a slight modification was used to determine inhibition of glucose oxidase level. 0.05 ml glucose solution of 20, 40, 60, 80& 100 mg/dl was added wit h 0.05ml of extract in to different test tubes and incubated in dark for 4hrs. Then 5 ml of glucose oxidase enzyme was added in all test tubes and kept for 30 minutes in dark at RT. Res ults were spectrophotometrically (Thermo scientific Multiscan Go) recorded at 546nm. Concentration of glucose was meas ured as following Ak Concentration of glucose= ?????? - X C Au Where Ak = Absorbance of known (standard glucose) ; Au = Absorbance of unknown (extracts of plants), C = conc entration of standard glucose. Statistical Analysis 2 All samples were tested and analyzed in triplicate. Results were calculated as the mean ? SD (standard deviation) fo r each sample. Statistical analysis was done with one way analysis of variance using Graph pad Prism, Version 4.0 (Graph Pad Software, San Diego, CA, USA). A significant differ ence was judged to exist at a level of P < 0.05.
Our study reveals the uses of glucose utilization u sing enzyme glucose oxidase method in a complex mixture or an e xtract because of its specificity. The phytochemical analy ses of five bamboo species of North Bengal region were assessed which revealed their hypoglycemic activity. The results indicate the presence of active constituents in the solvents ext racted from medicinal plants material. Special consideration to these effective therapeutic plants will lead us to obtain innovative medicines in controlling diabetes mellitus. 3.1 In vitro Hypoglycemic activity at different pH Figure I, II & III display anti hyperglycemic activity of the different plants at the different pH 2, 7 and 9 res pectively. At pH 7 and 9, extracts showed considerable anti-hyper glycemic activity in comparison to pH 2. At a concentration of 80 ?g/ml, B. pallida and B. vulgaris at basic and neutral pH however B. balcooa showed the highest antihyperglycemic activity at acidic pH. The statistical analysis indicated signi ficant difference between hypoglycaemic activity of plant extracts at P< 0.05. One of the possible reasons for these plan ts could be its adenosine deaminase inhibitory activity, hence reduces glucose levels in hyperglycemic patients as suggest ed in some other plants studies 13 . Previous studies revealed that B. vulgaris repairs damaged ? -cells, increases insulin levels, and also enhance the sensitivity of insulin. It might inhibi t glucose 29 oxidase and therefore glucose absorption and also s uppresses the activity of disaccharides in the intestine 15 . Since these plants are found to have good antihyperglycemic act ivity in vitro . Further, in vivo investigations are needed to prove their anti-hyperglycemic activity. Even more comprehensiv e chemical and pharmacological studies are needed to exploit their ethano-medicinal relevance.
B. vulgaris, B. pallida, B. balcooa, D. hamiltonii and D. sikkimensis provided the imprint of being noticeable candidates for drug targets for diabetes. This may perhaps be the first strategy using in vitro methods to verify their anti- hyperglycemic activity.
Authors acknowledge Dr. TL Shantha, Director, resea rch centre, Dr. MB Nagaveni, Dean of Life Sciences, MLAC W for providing the facilities and interpretation softwar e. Authors would like to thank Dr. AK Goyal, NBU, Siliguri, for helping in collection of samples.
1. Middha SK, Mittal Y, Usha T, Kumar D, Vashist L & Nagaveni MB, Phyto-mellitus: A phyto chemical database for diabetes. Bioinformation , 4 (2009)78-79. PMID:20198174 2. Middha SK, Bhattacharjee B, Saini D, Baliga MS, Nagaveni MB & Usha T, Protective role of Trigonella foenum graceum extract against oxidative stress in hyperglycemic rats. Eur Rev Med Pharmacol Sci , 15 (2011) 427?435. PMID:21608438 3. Singh R, Singh MK, Chandra LR, Bhat D, Arora MS, Nailwal T & Pande V, In vitro Antioxidant and free radical scavenging activity of Macrotyloma uniflorum (Gahat dal) from Kumauni region. Int J Fundam Appl Sci, 1 (2012) 7?10. 4. Goyal AK, Middha SK & Sen A, Evaluation of the DPPH radical scavenging activity, total phenols and antioxidant activities in Indian wild Bambusa vulgaris ?Vittata? methanolic leaf extract. J Nat Pharm, 1 (2010) 40-45. 5. Goyal AK, Middha SK, Usha T, Chatterjee S, Bothra AK, Nagaveni MB & Sen A. Bambooinfoline: a database for North Bengal bamboos. Bioinformation, 5 (2010) 184-85. 6. Goyal AK, Ghosh PK, Dubey AK & Sen A. Inventorying bamboo biodiversity of North Bengal: A Case Study. Int J Fundam Appl Sci, 1 (2012) 5-8. 7. Ranjbar SH, Larijani B, Abdollahi M. A systematic review of Iranian medicinal plants useful in diabetes mellitus. Arch Med. Sci , 4 (2008) 285?292. 8. Aggarwal N & Shishu, A Review of Recent Investigations on Medicinal Herbs Possessing Anti diabetic Properties. J Nutrition Disorder Ther, 1 (2011)102. 9. Khan B, Arayne SA, Nazs S & Mukhtar N, Hypoglycemic activity of aqueous extract of some indigenous plants. Pak J Pharm Sci , 18 (2005) 62-64. PMID : 16431387 10. Goyal AK, Middha SK & Sen A, In vitro antioxidative profiling of different fractions of Dendrocalamus strictus (Roxb.) Nees leaf extracts. Free Rad. Antiox, 1 (2011) 42-48. 11. Trinder P, Determination of glucose in blood using glucose oxidase with an alternative oxygen acceptor. Annals Clin Biochem, 6 (1969) 24-27. 12. Bopp A, De-Bona KS, Bell? LP, Moresco RN & Moretto MB. Syzygium cumini inhibits adenosine deaminase activity and reduces glucose levels in hyperglycemic patients. Fundam Clin Pharm , 23 (2009) 501?507. PMID: 19709327 13. Chaturvedi P, Antidiabetic potentials of Momordica charantia : multiple mechanisms behind the effects. J Med Food, 15 (2012) 101- 107. PMID:22191631 14. Khan V, Najmi AK, Akhtar M, Agil M, Mujeeb M & Pillai KK, Evaluation of Traditional Indian Antidiabetic Medicinal Plants for Human Pancreatic Amylase Inhibitory Effect In Vitro A pharmacological appraisal of medicinal plants with antidiabetic potential. J Pharm Bioallied Sci , 4 (2012)27-42. (PMID:22368396).
Plants, a therapeutic source are a prehistoric know ledge 1 . Past investigations have indicated that the consumption of natural antioxidants is related with reduced risks of numer ous ailments like diabetes 2 . Freshly, there is an increasing attention in finding natural antioxidants to substitute manmade ones 3,4 . India has a vast biodiversity due to its topographi cal and climatic circumstances. In India about 139 species of bamboos are encountered belonging to 36 genera 5 . North east region holds two third of the countries bamboo reserve wit h 58 species included in 10 genera. Ethano-pharmacological surve y studies in this region have revealed a large number of plan t species are traditionally used in North east and surround regio ns for many ailments 6 . There are a few or almost none studies on these plants that have been recorded. Also, these plants were recommended by the natural healers for treatments o f diabetic symptoms and its complications 1,7,8 . Numerous in vivo methods are there to investigate anti- hyperglycemic activity of these herbs. But there are rare specific studies 9 to verify their in vitro anti-hyperglycemic activity. The current contemporary practice could b e used to diminish animal sacrifices in the preliminary scree ning of anti- hyperglycemic activity of plant species. Neither pr evious reports provide any demonstration on in vitro anti-hyper glycemic studies of these ethnic plants nor on whic h medium or pH does the plant illustrate remedial activity. The refore, this study is premeditated to witness the inhibition of glucose oxidase, a marker of anti-hyperglycemic activity of therapeutic plants used in north east traditional system in Ind ia.