Development of a polyherbal formulation FMST and evaluation for antidiabetic activity in alloxan induced diabetic rats

 

Alimuddin Saifi¹*, Rajani Chauhan², Jaya Dwivedi³

¹Dept. of Pharmacognosy, NKBR College of Pharmacy and Research Centre, Meerut (U.P.)

²Dept. of Pharmacy, Banasthali University, Rajasthan

³Dept. of Chemistry, Banasthali University, Rajasthan

 

ABSTRACT:

Background: Medicinal plants have curative properties due to the presence of various complex chemical substance of different composition, which are found as secondary plant metabolites in one or more parts of these plants. Herbal formulation treatment can significantly alter the pattern of glucose tolerance in normal and diabetic rats. It is possible that herbal formulation may act through both, pancreatic and extra pancreatic mechanism(s). In present study the polyherbal formulation (FMST) was developed by mixing the hydroalcoholic extracts of the stem barks of Ficus bengalensis (FBE), fruits of Momordica charantia (MCE) and seeds of Trigonella foenum graecum (TGE) and Syzygium cumini (SCE) in optimized ratio. The aim of this work was to establish the dose response relationship of the individual and combined herbal extracts. Further it was aimed to optimize the dose of the formulation to produce the required effect.  The antidiabetic activity was determined in normal and diabetic rats respectively. In the hypoglycemic study of individual extracts, the optimum doses were found to be 120, 300, 1000 and 500 mg/kg b.w, p.o/day, of extracts of FB, MC, TG and SC respectively. The optimized doses were further evaluated for antidiabetic activity in alloxan induced diabetic rats. Based on the above optimized doses, a polyherbal formulation was prepared by mixing the extracts in the ratio of 1.2:3:10:5.

Materials and method: Albino Wister male rats of weighing between 150 to 200 gms used for the study. Diabetes was induced by injecting alloxan (120 mg/kg, i.p.). Group I served as normal control, Group II served as diabetic control, Group III served as standard control and treated by Tolbutamide 100 mg/kg p.o. Group IV, V and VI served as diabetic rats treated with FMST at different dosage of 200 mg/kg (FMST-2), 400 mg/kg (FMST-4) and 600 mg/kg (FMST-6) for the purpose of dose optimization and to find out the most effective and safer dose. All the treatments were given for 21 days. At the end of study on 21 day over night fasted rats were sacrificed and blood was collected to determine fasting blood glucose and biochemical findings.

Result: The dose response relationship of the formulation was observed as per the evaluation of antidiabetic activity. The combined herbal extract exhibited synergistic effect, and was better than any of the extracts. The initiation of the effect was early and the duration was increased with the combination. Diabetic rats treated with FMST-2, FMST-4 and FMST-6 significantly (P<0.01) reduced fasting blood glucose and normalize the lipid profile, renal profile and hepatic profile. Improvement in the histopathology of pancreas and liver of FMST-4 treated rats confirmed its protective role in alloxan induced diabetes. Conclusion: It can be concluded that FMST-2, FMST-4 and FMST-6 possess antidiabetic activity. FMST-4 was found to be optimum and may be beneficial improving complications associated with diabetes mellitus. From the above, it may be concluded that polyherbal formulation exhibited superior desired activity because of their combined individual activities. The study provides scientific support for their claimed activity in Ayurveda.

 

 

INTRODUCTION:

In the recent years there is a great interest towards Ayurvedic forms of medication not only for diabetes but also for other disease like arthritis and cancer as the danger and shortcoming of modern medicine have started getting more apparent. Majority of Ayurvedic formulation are prepared from herb and many herbal formulation are in market and are immensely used by diabetic patients on the advice of physicians.¹ Herbal formulation treatment can significantly alter the pattern of glucose tolerance in normal and diabetic rats. It is possible that herbal formulation may act through both, pancreatic and extra pancreatic mechanism(s).² It is evident that there are many herbal formulations of varying potency since these preparation act by different mechanism, it is theoretically possible that different combination of these extract will do better job in reducing blood glucose. In the traditional system of plant medicine it is usual to use plant formulation and combined extract of plant are used as a drug of choice rather than individual ones.³

 

The bark of the Ficus bengalensis has been reported to possess hypocholesterolaemic and hypolipidaemic effects. A glycoside, leucopelargonidin was also isolated from the bark of Ficus bengalensis and its antidiabetic effects have been reported.⁴ It acts by the mechanism of rising serum insulin in diabetic rats.⁵ Momordica charantia enhances insulin secretion and increases the number of pancreatic B-cells in the islets of Langerhans and increases the glucose uptake in liver via promoting glucose-6- phosphate dehydrogenase and declining glucose-6- phosphatase activities.⁶ Fenugreek (Trigonella foenum-graecum) has primarily been described as an antihyperglycemic herb in humans as well as in laboratory animals.^{7, 8} It’s cholesterol-reducing effect is also well established.⁹ Syzygium cumini show significant antihyperglycemic activity in mild diabetic rats which have functioning pancreatic β cells indicating that it may modulate insulin release and an increase in insulin level. Results in concordance with earlier reports of S. cumini was found to increase insulin secretion.¹⁰ S. cumini has been reported to work as antidiabetic by enhancing cathepsin B activity thereby increasing the proteolytic conversion of proinsulin to insulin¹¹ and inhibits insulinase activity of liver and kidney.¹²

 

The present paper deals with development of polyherbal formulation (FMST) by mixing the hydroalcoholic extracts of the stem barks of Ficus bengalensis, fruits of Momordica charantia and seeds of Trigonella foenum graecumand Syzygium cumini in optimized ratio and evaluation of anti-diabetic activity. The Poly Herbal Formulation was prepared in three different strengths for the purpose of dose optimization and to find out the most effective and safer dose.

 

MATERIAL AND METHODS:

Plant materials

The fruits of Momordica charantia and seeds of Trigonella foenum graecum and Syzygium cumini were purchased from local market Hapur (U.P.). The stem bark of F. bengalensis Linn. was collected in the month of august from Delhi road, Hapur. The specimen was given for authantification in Raw Material and Laboratory of National Institute of Science Communication and Information Resources (NISCAIR), New Delhi (voucher no. NISCAIR/Consult/RHMD/-2010-11/1620/218).

 

The crude drugs were washed and dried in an electric hot air oven at a temperature 40^o C.

 

Extraction of plant materials

The stem barks of Ficus bengalensis, seeds of Syzygium cumini and Trigonella foenum graecum and fruits of Momordica charantia are dried in a hot air oven at temperature of 40⁰C. Dried plant materials were coarsely powdered and defatted with petroleum ether by soxhlet apparatus. Defatted drug than exhaustively extracted with 70% v/v hydro alcoholic solution using soxhlet apparatus. The extract was concentrated under reduced pressure.

 

Standardization of plant drugs and extracts

The morphological and microscopical studies, ash value, extractive value in different solvents, qualitative heavy metal analysis of the extract and test for microbial contamination were also done for the purity of drug. The Phytochemical screening of the extracts were carried out for the presence of Alkaloids, Proteins and Amino acids, Carbohydrates, Flavonoids, Phenolic group, Glycosides, Saponins, Tannins, Steroids, Triterpinoids. The heavy metal analysis was performed as per guidelines of Indian Pharmacopoeia. Microbial count included total viable aerobic count, total yeast and mould, E. coli, S. typhi, P. aeruginosa and S. aureus count have been determined.^13-20 The results of standardization, antidiabetic activity and histopathological studies of all the above mentioned drugs are published in previous papers.^13-16

 

Development of Poly Herbal Formulation:

The Poly Herbal Formulation was prepared in three different strengths, 200 mg/kg b.w, p.o/day  (FMST-2), 400 mg/kg b.w, p.o/day (FMST-4), 600 mg/kg b.w, p.o/day (FMST-6) for the purpose of dose optimization and to find out the most effective and safer dose.

The following formulations were prepared:

 

Table1: Formulation 1 (FMST-2)

Ingredients	Quantity (mg)
Ficus bengalensis (bark extract)	12.5 mg
Momordica charantia (fruit extract)	31.5 mg
Syzygium cumini(seed extract)	52.08 mg
Trigonella foenum graecum(seed extract)	104.16 mg

 

Table2: Formulation 2 (FMST-4)

Ingredients	Quantity (mg)
Ficus bengalensis (bark extract)	25 mg
Momordica charantia (fruit extract)	62.5 mg
Syzygium cumini(seed extract)	104.16 mg
Trigonella foenum graecum(seed extract)	208 mg

 

Table3: Formulation 3 (FMST-6)

Ingredients	Quantity (mg)
Ficus bengalensis (bark extract)	37.5 mg
Momordica charantia (fruit extract)	93.75 mg
Syzygium cumini(seed extract)	156.25 mg
Trigonella foenum graecum(seed extract)	312.5 mg

 

Animals

Albino wistar male rats of weighing between 150 to 200 gms were procured from Indian Veterinary Research Institute Bareilly U.P. (IVRI).The animals were housed under standard conditions of temperature (25 ± 2⁰C) and relative humidity (30-70%) with a 12:12 light-dark cycle and acclimatized in the animal house facility of the department under ambient condition of Siddhartha Institute of pharmacy, Dehradun (CPCSEA Approval no. 1435/PO/a/11/CPCSEA). The animals were fed with standard diet (Amrut Rat Feed, India) and water ad libitum. The Institutional Animal Ethics Committee approved all the experimental protocols with approval no. SIP/IAEC/12/Polyherbal.

 

Acute Toxicity studies

The acute oral toxicity study was carried out as per the guidelines set by Organization for Economic Co-operation and Development (OECD), received draft guidelines 420, received from committee for the purpose of control and supervision of experiments on Animals (CPCSEA), Ministry of Social Justice and Empowerment, Government of India. A total of six female albino rats were used for the study. The Poly Herbal Formulation (FMST) was administered in a single dose of 3000 mg/kg by gavage. All animals were observed individually after dosing during first 30 minutes, periodically during the first 24 hours and daily for 14 days. ¹⁷

 

Induction of Diabetes:

The albino rats were allowed to fast overnight prior to experimentation and rendered diabetic by injection a single dose of Alloxan 120 mg/kg body weight (Manufactured by Loba Chemie Company) administered as a 0.9% w/v in saline solution by I.P. route. It produces diabetes by selective necrosis of b - cells of islets of langerhans of pancreas. Since alloxan could evoke fatal hypoglycemia as a result of massive insulin release, rats received 20% of glucose solution for first 6 hr then simple tape water was given. The rats were then kept for next 24 hr with free access of 5 % glucose solution to prevent hypoglycemia.¹⁸ After 48 hrs of injection of Alloxan, Blood glucose level was measured for the evidence of diabetes by using commercially available kit “ACCU-CHEK ACTIVE” Glucometer from Roche Diagnostics GmbH, Germany. The rats which showed blood glucose level more than 200 mg/dl were considered as diabetic. The animals with sugar level more than 200mg/dl were selected. Animals were maintained for 72 hrs in diabetic condition for well establishment of diabetes.

 

Experimental design

The Poly Herbal Formulations (FMST) were suspended in 2% acacia solution and the given by oral route using a catheter. Tolbutamide 100mg/kg was used as a standard drug.¹⁹

Animals were divided into six groups of six each.

Group-1:  Healthy normal animals received only the water served as Normal control (NC).

Group-2: Untreated, alloxan induced diabetic animals served as a Diabetic control group (DC) also received water.

Group-3: The Reference Standard group (STD) was treated with Tolbutamide 100 mg/kg b.wt., p.o.

Group-4: Diabetic animals treated with FMST-2.

Group-5: Diabetic animals treated with FMST-4.

Group-6: Diabetic animals treated with FMST-6.

 

Blood samples were collected by retro-orbital plexus puncture method and blood glucose levels were estimated using an electronic glucometer (ACCU-CHEK ACTIVE” Glucometer from Roche Diagnostics GmbH, Germany). Blood samples were drown at weekly intervals till the end of study (i.e. 3 weeks). Blood glucose estimation was done on day 0, 7, 14 and 21 of the study. On day 21, blood was collected by cardiac puncture under mild ether anesthesia from overnight fasted rats and fasting blood sugar²⁰ was estimated. Serum was separated and analyzed for serum cholesterol,²¹ serum triglycerides,²² serum HDL,²³ serum LDL,²⁴ serum creatinine,²⁵ serum urea,²⁶serum alkaline phosphatase (ALP),²⁷ bilirubin, serum glutamate oxalate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT)²⁸ were estimated.

 

Statistical analysis

All values of blood sugar and biochemical estimations were expressed as mean ± Standard error means (S.E.M.) and analysed for ANOVA and post test TUKEY-One Way Analysis of Variance. Differences between groups were considered at P<0.01 levels. 

Histopathology of isolated liver and pancreas:

Small pieces of liver and pancreas tissues were collected in 10% formalin for proper fixation. Tissues were fixed in Bouin’s fixative (without acetic acid) for histopathological studies. Sections of tissues (6 microns in thickness) were stained with haematoxylin and eosin (H and E) for histological examination.²⁹ The photomicrographs of histological studies are presented in fig. 4 and 5.

 

RESULTS:

Acute Toxicity studies

FMST treated rats showed no discernible behavioral changes given by oral route. No mortality was observed when it was administered orally at a high dose (3g/kg bw) which is higher than effective antihyperglycemic dose during observation period. Closely observed for 24 hours for any mortality and up to 14 days for any delayed toxic effects on behavioral activities.

 

Antidiabetic activity

Table 4- Effect of 3-week treatment with standard drug and FMST on blood glucose level after alloxan induced diabetic rats

* P<0.01 (Tukey test) significant when treated with Normal control

# P<0.01 (Tukey test) significant when treated with Diabetic control

 

 

 

Fig.1: Graphical representation of effect of FMST on blood glucose after 7 days treatment

 

 

Fig.2: Graphical representation of effect of FMST on blood glucose after 14 days treatment

 

 

Fig.3: Graphical representation of effect of FMST on blood glucose after 21 days treatment

 

Table 5- Effect of FMSTon serum profile in alloxan induced diabetic albino rats after 21 days treatment

* P<0.01 (Tukey test) significant when treated with Normal control

 # P<0.01 (Tukey test) significant when treated with Diabetic control

 

 

Histopathology

Photomicrographs (Fig.4) shows normal acini and normal cellular population in the islets of langerhans in pancreas of normal control and lesions in diabetic rats which maintained significantly after treatment by standard drug and FMST-4 up to normal.

 

 

Photomicrographs (Fig.5) shows normal hepatocytes and lesions in diabetic rats which maintained significantly after treatment by standard drug and FMST-4.

 

Fig. 4: Photomicrograph of rat pancreas stained by haematoxylin and eosin of normal control (A) diabetic control (B) standard (tolbutamide) treated (C) FMST-2 treated (D) FMST-4 treated (E) FMST-6 treated (F)

 

Fig. 5: Photomicrograph of rat liver stained by haematoxylin and eosin of normal control (A) diabetic control (B) standard (tolbutamide) treated (C) FMST-2 treated (D) FMST-4 treated (E) FMST-6 treated (F)

 

DISCUSSION:

Medicinal plants are the potential source of bioactive agents and gaining acceptability worldwide. The ethnobotanical prospect can help for the development of drugs to treat human diseases like diabetes. Safe, effective and inexpensive indigenous remedies are gaining popularity equally among the people of both the urban and rural areas, especially in developing countries like India.³⁰

 

Alloxan induced diabetes has been commonly utilized as an animal model to study diabetes in experimental animals. Alloxan exerts it’s diabetogenic actions when administered intravenously, intraperitoneally or subcutaneously. The action of alloxan in the pancreas is preceded by its rapid uptake by the insulin-secreted beta cells.³¹ The cytotoxic action of alloxan is mediated by reactive oxygen species which leads to rapid destruction of beta cells, thereby reducing levels of insulin and increasing the blood glucose.^{32, 33} However there is possibility of survival of a few beta cells and this has been proved by several workers who observed antihyperglycemic activity with oral hypoglycemic agents in alloxan induced diabetes mellitus.^{34, 35}

 

The polyherbal formulations FMST-2, FMST-4 and FMST-6 have been shown marked glucose lowering effect as shown in Table No. 4. FMST-6 significantly reduced blood glucose from 352.3 ± 6.216 to 156.3 ± 14.53 which was maximum than FMST-2 and FMST-4 after 21 days treatment. The histology of pancreas photomicrographs (Fig.4) shows normal acini and normal cellular population in the islets of langerhans in pancreas of normal control and lesions in diabetic rats which maintained significantly after treatment by standard drug FMST-4 and FMST-6 up to normal. FMST-6 had shown the best result. The histology of liver in photomicrographs (Fig.5) shows normal hepatocytes and lesions in diabetic rats which maintained significantly after treatment by standard drug and FMST-4, but FMST-6 shows lesions in diabetic rats which are not maintained significantly after treatment this indicates that high dose of the formulation (FMST-6) may be toxic to the liver. FMST-4 maintained hepatocytes up to the normal. So the result reveals that FMST-4 can be a safe acceptable and effective alternative or adjuvant to the conventional oral hypoglycemic agents.

 

There was a significant increase in β cell count in the FMST-4 treated group. Significant improvement in renal function and lipid profile could be secondary to reduction in blood sugar level.

 

The anti-hyperglycemic effect of FMST-2, FMST-4 and FMST-6 on the blood sugar levels of diabetic rat is shown in Table 4 and Fig. 1, 2 and 3.

 

Serum cholesterol, serum triglycerides, serum, serum LDL, serum creatinine, serum urea, serum alkaline phosphatase (ALP), bilirubin, serum glutamate oxalate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT) were significantly decreased by Tolbutamide and FMST-2, FMST-4 and FMST-6. HDL and albumin levels were significantly increased by Tolbutamide and FMST-2, FMST-4 and FMST-6. Effects on serum profile are shown in Table 5.

 

CONCLUSION

Thus our study proves the beneficial effects of polyherbal formulation FMST-4 in the management of diabetes and its associated complications. Our finding supports the long term use of the FMST-4 per day for better control of blood glucose and restorations of diabetes associated changes. After completing the preclinical studies the herbal product FMST-4 needs to be tried on human diabetic patients to ascertain its efficacy and safety.

 

ACKNOWLEDGEMENT:

The author wish to thank the Director, Siddhartha Institute of Pharmacy, Dehradun, for providing animal experimentation facilities and A.K. Diagnostics Services Laboratory Networks, New Delhi.

 

REFERENCES:

1.    Katiyar DK, Mehrotra Anju, Pant KK, Ali Basheer, Study of hypoglycemic effects and antioxidant potential of polyherbal formulation in rats, Biomedical Research, 2012, 23 (3), 444- 449.

2.       Mandlik RV, Desai SK, Naik SR, Antidiabetic activity of a polyherbal formulation (DRF/AY/5001), Indian Journal of Experimental Biology, 2008, 46, 599-606.

3.       Kumar Jaya. Herbal medicine for Type 2 diabetes. International Journal of Diabetes in Developing Countries, 2010, 30, 111-112.

4.       Gayathri M. and Kannabiran K., Antidaibetic and ameliorative potential of Ficus bengalensis bark extract in streptozotocin induced diabetic rats. Indian Journal of Clinical Biochemistry, 2008, 23(4), 394-400.

5.       Cherian S and Augusti KT, Antidiabetic effect of a glycoside leucopelargonidin isolated from Ficus bengalensis Linn. Indian Journal of Experimental Biology 1993, 31(1), 26-29.

6.       Abdollahi M, Zuki ABZ, Goh YM¹, Rezaeizadeh A and Noordin MM, The effects of Momordica charantia on the liver in streptozotocin-induced diabetes in neonatal rats, African Journal of Biotechnology 2010, 9(31), 5004-5012.

7.       Bordia, A, Verma SK, Srivastava KC. Effect of ginger (Zingiber officinale Rosc.) and fenugreek (Trigonella foenum-graecum L.) on blood lipids, blood sugar and platelet aggregation inpatients with coronary artery disease. Prostaglandins Leukot. ssent. Fatty Acids, 1997,56, 379–384.

8.       Sharma RD, Raghuram TC, Rao NS, Effect of fenugreek seeds on blood glucose and serum lipids in type I diabetes. Eur J Clin Nutr 1990, 44,301–6.

9.       Sharma RD. Hypocholester olemic activity of fenugreek (T. foenum- graecum). An experimental study in rats. Nutr Rep Int 1984, 30, 221– 31.

10.    Sharma SB, Nasir A, Prabhu KM, Murthy PS. Antihyperglycemic effect of the fruit-pulp of Eugenia jambolana in experimental diabetes mellitus. J Ethnopharmacol 2006, 104(3), 367–373.

11.    Bansal R, Ahmad N, Kidwai JR. Effects of oral administration of Eugenia jambolana seeds and chloropropomide on blood  glucose level and pancreatic cathepsin B in rat. Indian J Biochem Biophy, 1981, 18, 377–382.

12.    Achrekar S, Kaklij GS, Pote MS, Kelkar SM. Hypoglycemic activity of Eugenia jambolana and Ficus bengalensis: mechanism of action. In Vivo, 1991, 5(2), 143–147.

13.    Alimuddin Saifi, KP Namdeo, Rajani Chauhan And Jaya Dwivedi, Standardization and evaluation for antidiabetic activity of Ficus bengalensis linn. Stem bark, Indo American Journal of Pharmaceutical Research, 2014, 4 (04), 1960-67.

 

14.     Alimuddin Saifi, KP Namdeo, Rajani Chauhan And Jaya Dwivedi, Evaluation of pharmacognostical, phytochemical and antidiabetic activity fruits of Momordica charantia linn, Asian J Pharm Clin Res, 2014, 7(3), 152-156.

 

15.     Alimuddin Saifi, Rajani Chauhan and Jaya Dwivedi, Assessment of the antidiabetic activity of Syzygium cumini (Linn.) Skeels in alloxan induced diabetic rats, Research Journal of Pharmacology and Pharmacodynamics. 2016, 8(3), 91-96.

16.     Alimuddin Saifi, Rajani Chauhan and Jaya Dwivedi, Evaluation of antidiabetic activity of hydro-alcoholic extract of Trigonella foenum-graecum Linn. in alloxan induced diabetic rats, Asian Journal of Pharmacology and Toxicology, 2016, 04(15), 26-32.

17.    Revised Document (Dec-2001) OECD/OCDE 420. Guidelines on Acute Oral Toxicity. OECD Guidelines for the testing of chemicals, 1-14.

18.    Sangameswaran, B, Singh R, Balakrishnan BR, Jayakar B. Anti-hyperglycemic and anti-hyperlipidemic effects of Ipomoea reniformis Chois on alloxan induced diabetic rats. Journal of Research and Education in Indian Medicine 2007, 13(4)  27-30.

19.    Hemlatha S, AyyappanT, Shanmugam S, Nagavalli D, Shrivijaya Kirubha T, Evaluation of antidiabetic and anti diuretic activity of polyherbal formulation. Indian Journal of Traditional Knowledge, 2006, 5(4), 468-470.

20.    Giordano BP, Thrash W, Hollenbough L, Dube WP, Hodges C, Swain A, Banion CR, Klingensmith GJ. Performance of seven blood glucose testing systems at high altitude. Diabetic education 1989, 15, 444-448.

21.    Roeschlau P, Bernt E, Gruber W. Enzymatic determination of total cholesterol in serum. Zeitschrift fur Klinische Chemie und Klinische Biochemi, 1974, 12, 226.

22.    Muller PH, Schmulling RM, Liebich HM, Eggstein M. A fully enzymatic triglycerides determination. Journal of Clinical Chemistry and Clinical Biochemistry, 1977, 15, 457-464.

23.    Allain CC, Poon LS, Chan CS, Richmond W, Fu PC. Enzymatic determination of total serum cholesterol. Clinical Chemistry, 1974, 20, 470-475.

24.    Friedwald WT, Levy RI, Fredrickson DS. Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without use of preparative ultracentrifuge. Clinical Chemistry, 1972, 18, 499-502.

25.    Bowers LD. Kinetic serum creatinine assays I. the role of various factors in determining specificity. Clinical Chemistry, 1980, 26, 551-554.

26.    Wilson BW. Automatic estimation of urea using urease and alkaline phenol. Clinical Chemistry, 1966, 12, 360-368.

27.    Sasaki M. A new ultramicro method for the determination of serum alkaline phosphatase. Use of Berthelot’s reaction for estimation of phenol released by enzymatic activity. Igaku To Seibutsugaku, 1966, 70, 208-214.

28.    Henry RJ, Cannon DC, Winkelman DC. Enzymes, in. Clinical Chemistry: principles and techniques (Harper and Row NY) 1974, 842.

29.    Ramnik Sood, Concise book of Medical Laboratory Technology, Methods and Interpretations, Jaypee Brothers Medical Publishers(P) Ttd, New Delhi, 2009, 972-992.

30.    Katewa SS, BL Chaudhary and A Jain, 2004. Folk herbal medicine from tribal areas of Rajasthan, India. J. Ethnopharmacol., 92, 41-46.

31.    Szudelski T. The mechanism of alloxan and streptozotocin action in B cells of the rat pancreas. Physiol. Res., 2001, 50, 536-546.

32.    Cakici IC, B Hurmoglu, N Tuncton, AI Kanzik and B Sener. Hypoglycemic effect of Momordica charantia extract in normoglycemic mice. J. Ethnopharmacol., 1994, 44, 117-121.

33.    Abdel-Barry JA, IA Abdel-Hassan and MHH Al-Haliem. Hypoglycemic and Antihyperglycemic effects of Trigonella foenum-graecum leaf in normal and alloxan induced diabetic reats. J. Ethnopharmacol., 1997, 58, 149-155.

34.    Cherain S and KT Augusti. To study the antidiabetic effects of a glycoside of leucopelargonidin isolated from Ficus bengalensis Linn. Indian J. Exp. Biol., 1993, 31, 26-29.

35.    Kumari K, BC Mathew and KT Augusti. Antidiabetic and hypolipidemic effects of S-methyl cystine sulfoxide isolated from Allium cepa Linn. Indian J. Biochem., 1995, 32, 49-54.

 

 

 

 

Received on 20.10.2016       Accepted on 27.11.2016     

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