ISSN 2229-5054

International Journal of Drug Formulation & Research Jan-Feb. 2011, Vol. 2 (1)

International Journal of Drug Formulation & Research

ANTI DIABETIC ACTIVITY OF TRADITIONAL HERBAL FORMULATION

Abdullah khan*1, Sadat ali2, Jameel ahmad3
1) Singhania University, Pacheri baru Rajastan! K.C.T College of Pharmacy GULBARGA 585104 (Karnataka) INDIA 2) Luqman College of Pharmacy, Old Jewargi Road, GULBARGA 585102 (Karnataka) INDIA 3) H.K.E.S'S College of Pharmacy, M.R Medical College Campus GULBARGA 585105 (Karnataka) INDIA

Abstract:
An indigenious herbal formulation containing Methi (fenugreek), Sesamum indicum (black sesame seed), Acacia catechu (Katha safeed), Azadirachta indica (Neem leaves), Momordica charantia (karela) was evaluated for hypoglycaemic activity on adult wister albino rats each using normo glycemic, glucose loaded and alloxan induced hypoglycemic rats. Glibenclamide (2.5mg/kg) was used as reference standard for activity comparision. The formulation showed promising results that is comparable to that of the reference standard glibenclamide. The present work justifies the use of the herbal formulation in the traditional practice for the treatment of diabetes.

Keywords: Glibenclamide, Hyperglycaemic, Normoglycaemic, Oral glucose tolerance test (OGTT), Traditional herbal
formulation.

Introductions: Diabetes mellitus is a chronic disease caused by an absolute or relative lack of insulin and or reduced insulin activity, which results in hypoglycemia and abnormalities in carbohydrate, protein and fat metabolism. At least 171 million people worldwide have diabetes, this figure is

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likely to be more than double by 2030 and around 3.2 million deaths every year are attributable to complications of diabetes at the rate of six deaths every minute1. Search for antidiabetic factor in plants remains a potential area of investigation. In accordance with the recommendations of the WHO expert committee on diabetes mellitus, an investigation of antihyperglycaemic agents of plants origin used in traditional medicine seems important. Many herbs and plant products have been shown to have antihyperglycaemic action2. In the present study, an indigenous herbal formulation containing Methi (fenugreek), Sesamum indicum (Black sesame), Acacia catechu (katha safeed), Azadirachta indica (neem leaves) and Momordica charantia (karela)3-6. which claims to have the potential in the treatment of wounds, burns, fistula, inflammation etc, was selected for evaluation of hypoglycaemic activity on adult wistar albino rats, each using normoglycaemic, glucose loaded and alloxan induced hypoglycaemic rats. Glibenclamide (2.5mg/kg) was used as reference standard for activity comparision. Materials and methods: Preparation of formulation: The formulation was obtained as a gift sample from a traditional practioner Mr. Abdullah khan, Hyderabad. Animals: Swiss albino mice (20-25 g) of either sex were used for acute toxicity study and adult wistar albino rats (150-200 g) of either sex were used for the antidiabetic evaluation. The animals were kept in standard polypropylene cages at room temperature of 34 2 C and at 60-65 % relative humidity during the experimental work. The institutional Animal Ethical Committee approved all the experimental protocols. Acute toxicity study:

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Albino mice of either sex, weighting 20-25 g and at 90 days age were used to determine the dose. Animals were fasted for 18 hours with water ad libitum and the test formulation was administered by oral route to pairs of mice in ascending and widely spaced doses. 2 ml/kg body weight, 4 ml/kg body weight, 8 ml/kg body weight, 16 ml/kg body weight and 20 ml/kg body weight. Animals were observed continuously for two hours and occasionally further four hours and finally over night mortality was recorded by Up and down method7. No mortality was observed even after administration of a dose of 20 ml/kg body weight. Thus based upon the studies, doses of 5 ml/kg body weight and 10 ml/kg body weight were selected for the assessment of antidiabetic activity. Using normoglycaemic rats: The method of Bhopale et al., 20068 was followed. The animals were fasted for 18hours but were allowed free access to water before and throughout the duration of experiment. At the end of the fasting period, taken as zero time (0 hour), blood was withdrawn from the tip of the tail of each rat under anaesthesia and the blood glucose was estimated with Senso card blood glucose meter. The normal rats were then divided into four groups of six animals each. Group- served as solvent control and received only vehicle (2 ml/kg) through oral route. Group- received glibenclamide (2.5mg/kg). Group- received 5ml/kg body weight of test formulation. Group-V received 10ml/kg body weight blood glucose levels were measured after 1, 2, 4 and 6 hour of administration of single dose of the test samples. The results are depicted in table 1. Oral glucose tolerance test (OGTT) in rats: The method of Badole et al. 20079 was followed. Fasted rats were divided into 4 groups of six rats each. Group- served as a control and received only vehicle (2ml/kg) through oral route. Group- received glibenclamide (2.5mg/kg). Group- received 5ml/kg body weight of test formulation. Group-V received 10ml/kg body weight. After 30 minute of treatment, rats of all groups were loaded orally with glucose (2gm/kg). Blood samples were collected before and at
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30, 90 and 150 minute after glucose administration as per the method described earlier. The results are depicted in table 2. Alloxan induced hyperglycaemic rats: The method of Dash et al., 2008 was followed10. The acclimatized animals were kept fasting for 24 hours with water ad libitum and injected intraperitonially a dose of 120mg/kg of alloxan mono hydrate in normal saline. After one hour, the animals were provided standard laboratory diet ad libitum. The blood glucose level was checked before alloxanisation and 24 hour after alloxanisation by withdrawing blood from the tip of the tail of each rat under anaesthesia. The blood glucose level was measured as above. Animals were considered diabetic when the blood glucose level was raised beyond 225mg/dl as suggested by Edwin et al, 200711. This condition was observed at the end of 48 hours after alloxanisation. The animals were segregated into four groups of six animals in each. Group- served as negative control and received vehicle (2ml/kg) through oral route.Group-II received glibenclamide (2.5mg/kg). Group-III, received 5 ml/kg and Group-V 10 ml/kg of test formulation.Blood glucose level was estimated at 0, 1, 2, 4,and 6 hours respectively after administration of single dose of test samples. The results are depicted in (Table3). Statistical Analysis: The data obtained was subjected to one way of analysis of variance (ANOVA) for determining the significant difference. The inter group significance was analysed using Dunnet,st test. AP-value<0.05 were considered to be significant. All the values were expressed as mean SEM. Results and discussion: The results of normoglycaemic study (Table 1) reveals that the test formulation exhibited significant reduction in blood glucose concentration as compare to control. It was observed that the test formulation at dose of 5ml/kg body weight and 10ml/kg body weight reduced 24.71%

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and 33.97% blood glucose levels respectively, where as glibenclamide showed 46.49% in rats after six hour treatment. The effect of test formulation on glucose tolerance test in normal rats is shown in table 2. At 30 minute after glucose administration the peak of blood glucose level increased rapidly from the fasting value and then subsequently decreased. Both the tested formulations (5ml and 10 ml/kg body weight) exhibited significant hypoglycaemic effect but glibenclamide and test formulation (10ml/kg) significantly depressed the peak of blood glucose level at 90 minute after glucose loading. In the anti-hyperglycaemic study (table3), the rise in blood glucose level was observed after 24 hour of alloxanisation to the animals. Single administration of the test formulations in diabetic rats showed significant reduction in blood glucose level, where as the test formulation (10ml/kg body weight) showed to maximum reduction in blood glucose level (62.12%) as compared to other formulation under test at the end of six hour. The results of test formulation (10ml/kg) are comparable to that of the reference standard glibenclamide. The exact biological active constituent(s) responsible for the hypoglycaemic effect are neither reported nor the extract mode of action of the hypoglycaemic activity was reported earlier, with the lone observation that it is used in traditional diabetic treatment. The results indicate that the polyherbal formulation is effective in decreasing the blood glucose level in normal and diabetic animals. Hence, justifying its use in traditional practice. However, further investigation is required to know the exact mechanism of hypoglycaemic action. Acknowledgement: The authors are greatful to Dr.M.G.Purohit, Professor Emeritus, Luqman College of Pharmacy, gulbarga for the guidance and encouragement in carrying out this work.

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Reference: 1. WHO (World Health Organization)Global strategy on diet, physical activity and healthwww.who.int/dietphysicalactivity/publications/facts/diabetes/en;Accssed May, 2007 2. Pal D.C.,Jain S.K., Tribal Medicine,(Naya Prakash, New Delhi; 2000) pp.246-247 3. K.R. Kirtikar and B.D. Basu, Indian Medicinal Plants, International Plants, Oriental Enterprises, Dehra Dun (2001). 4. W.C. Evans, Trease and Evans Pharmacognosy, 15th Edn, Saunders Publication, Edinburgh. 5. Hakeem Mohd. and Zahoorul Hasan, Selected Herbo minernal Drugs, First Edition,Islamic Organization for Medical Science of Kuwait (1989). 6. C.K. Kokate, A.P. Purohit and S.B. Gokhale, Text Book of Pharmacognosy, Nirala Prakashan, Pune, (2001). 7. M.N.Ghosh, Fundamentals of Experimental Pharmacology, 3rd Ed., Hilton and Company, Kolkata(2005). 8. Bhopal G.M., More S.M., Damame M.M. and Nanda R.K. Antihyperglycaemic activity of Phyllanthus reticulates leaves extract in normal and alloxan indused diabetic mice. Indian Drugs. 44(8):615-17 (2007). 9. Badole S., Patel N., Bodhankar S., Jain B. and Bhardwaj S., Antihyperglycaemic activity of aqueous extract of leaves of Cocculus birsutus (L) Diels in alloxan induced diabetic mice. Indian J.Pharmacol. 38: 49-53 (2006).

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10. Dash G.K., Bal S.K., Annapurna M. M. and Suresh P., Studies on the hypoglycaenic activity of Hemidesmus indicus r. Br. Roots. Phcog Mag. 4(16):221-25 (2008). 11. Edwin E., Sheeja E., Dhanabal S.P. and Suresh B., Antihyperglycemic activity of Passiflora mollissima Bailey, Indian J.of Pharmaceutical Sciences 69(4):570-571(2007).

ILLUSRATIONS:

Table 1. Effect of polyherbal formulation on the blood glucose level in normal rats Blood glucose concentration (mg /dl)(normoglycaemic study)
Time (h) after treatment
Group I II Treatment Control Glibenclamide Dose 2ml/kg b.w 2.5mg/kg b.w Fasting 95.831.48 95.51.95 1 96.661.10 2 97.161.05 4 96.831.12 6 97.160.99

84.002.68* 79.661.91** 65.001.58** 49.670.66** (11.91%) (16.41%) (19.68%) (47.49%) 91.831.75 82.16+0.92** 76.664.23** 72.664.71** (6.38%) (16.13%) (21.68%) (25.75%) 86.662.73 (11.14%) 80.831.31** (17.05%) 70.671.49** (27.35%) 63.160.13** (34.97%)

III

Test formulation

5ml/kg b.w

98.162.33

IV

Test formulation

10ml/kg b.w

97.661.21

Values are expressed as mean S.E. (n=6).All columns are significant using ANOVA.; *P<0.05, *P<0.01 when compared to control;

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Table 2. Effect of polyherbal formulation on oral glucose tolerance in normal rats

Blood glucose concentration (mg /dl) (oral glucose tolerance study)

Post treatment
Group I Treatment Control Dose 2ml/kg b.w Fasting 91.331.92 30 min. 129.331.64 90 min. 135.66 1.15 150 min. 149.66 4.32

II

Glibenclamide

2 .5mg/kg b.w

90.172.79

114.008.51

98.00 8.50 ** (13.91 %)

81.336.19 ** (28.40%)

III

Test formulation

5ml/kg b.w

93.341.78

39.004.87

121.00 9.75(12.85 %)

99.005.22** (28.57%)

IV

Test formulation

10ml/kg b.w

92.662.65

140.334.57

106.00 1.39 *(24.29 %)

86.673.02** (37.96%)

Values are expressed as mean S.E. (n=6). All columns are significant using ANOVA. ; *p<0.05, **p<0.01 when compared to control;

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Table 3. Effect of polyherbal formulation on the blood glucose level in alloxan induce diabetic rats

Blood glucose concentration (mg / dl)

Group I Treatment Control Dose 2 ml/kg b.w 0 hour 233.007.21 1 hour 238.195.61 2hour 245.555.17 4hour 251.54.65 6hour 254.663.33

II

Glibenclamide

2.5 mg/kg b.w

231.333.06

204.666.84*

177.52.79** (47.13%)

121.835.27**

82.006.30**

(11.47 %)

(23.16 %)

(64.27%)

III

Test formulation

5 ml/kg b.w

235.677.90 (1.90%)

231.167.98 (5.91%)

221.666.91

171.8313.5** (26.97%)

134.337.51** (42.81%)

IV

Test formulation

10 ml/kg b.w

240.336.75 (4.62%)

229.165.21

203.004.06** (15.13%) (33.28%)

160.005.92** (63.12%)

88.005.39**

Values are expressed as mean S.E.(n=6). All columns are significant using ANOVA. ; *P<0.05, **P<0.01 when compared to control;

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