Study of Antidiabetic activity of the stem bark extract of Holoptelea integrifolia (Roxb.)

 

Jyoti Sharma*, S.C. Sharma, Y.S. Sarangdevot, Bhupendra Vyas

B.N. College of Pharmacy, Udaipur (Rajasthan

 

 

ABSTRACT:

The plant Holoptelea integrifolia (Roxb.) belongs to Ulmaceae family. This species is used traditionally for the treatment of edema, diabetes, leprosy, skin diseases, intestinal disorders and piles. The present study of antidiabetic activity of the stem bark extract of Holoptelea integrifolia (Roxb.) was investigated in alloxan induced diabetic rats. The treatment was continue for 21 days. High mortality rate was seen in alloxan induced diabetic rats. The blood glucose level, body weight and lipid profile was estimated. The blood glucose level decreases gradually in the animals treated with the bark extract (250 mg/kg b.w.and 500 mg/kg b.w., orally), it was found to be statistically significant after 15 day treatment at dose level of 500 mg/kg.The prevention of loss of body weight in diabetic animals was found to be significant on day 15 in the animals of both treatment groups.

 

KEYWORDS: Holoptelea integrifolia; alloxan; blood glucose level; diabetes mellitus

 

INTRODUCTION:

Diabetes mellitus is a group of syndromes characterized by hyperglycemia; altered metabolism of lipids, carbohydrates, and proteins; and an increased risk of complications from vascular disease1. Diabetes is a major degenerative disease in the world today, affecting at least 15 million people and having complications which include hypertension, atherosclerosis and microcirculatory disorders. Diabetes mellitus is also associated with long-term complications, including retinopathy, nephropathy, neuropathy and angiopathy and several others2.

 

Holoptelea integrifolia (Roxb.) belonging to the family Ulmaceae. It is commonly known as Indian Elm tree. Holoptelea integrifolia is a large deciduous tree distributed throughout the greater part of India up to an altitude of 2,000 feet3. Ethnomedically used for the treatment of inflammation, acid gastritis, dyspepsia, flatulence, colic, intestinal worms, vomiting, wounds, vitiligo, leprosy, filariasis, diabetes, hemorrhoids, dysmenorrhoea and rheumatism4. It is an important pollen allergen of India5 and sensitizes almost 10% of the atopic population6. The bark and leaves are bitter, astringent, thermogenic, anti-inflammatory, digestive, carminative, laxative, depurative, anthelmintic and repulsive7-9.

 

Recently, the activities such as anti-inflammatory10,11, adaptogenic12 antidiarrhoeal13, anthelmintic14,15, wound healing16, antioxidant16,17, antimicrobial17-21, antitumor22 and antidiabetic23  activity have been reported in this plant.

 

MATERIALS AND METHODS:

Collection, identification and extraction of plant material:

Stem bark of Holoptelea integrifolia was collected from campus of B.N. College of pharmacy, in the month of June 2011. The tree was identified by Dr. J.S. Rathore, Professor in Botany B.N.P.G. College, Udaipur (Rajasthan).The stem bark was dried under shade and then powdered and stored in air tight container. Stem bark powder was subjected to extraction by maceration method using hydroalcoholic (50:50) solvent.

 

Animals:

The experimental animal albino rats were used in the study. Animals were housed in polypropylene cages at the room temperature and provided standard rodent feed and water ad libitum in a 12 hr light dark cycle.

 

Induction of diabetes:

Diabetes was induced by a single i.p. injection of 120 mg/kg Alloxan monohydrate24 (Sigma, Aldrich chemic) dissolved in sterile 0.9% saline. Rats were made to fast prior to alloxan administration.

 

Procedure:

The animals were divided into four groups of six animals each and were administered treatment according to the groups i.e. group1 (normal control), group 2 (diabetic control), group 3(diabetic test group 1 dose 250 mg/kg orally) and group 4 (diabetic test group 2 dose 500 mg/kg orally). Alloxan was administered in rats (120 mg/kg i.p.), after 6 hour 10% glucose solution was offered to animals to prevent hypoglycaemia. After 72 hour, blood glucose level was checked, to ensure development of hyperglycemia in groups 2, 3 and 4. From the fifth day the treatment was started, drugs were administered by oral route and the treatment was continue for 21 days. During the study period blood glucose level was determined again on days 0 (before treatment 4th day), 1, 3, 7 and 15 and 21day using glucometer. At the end of the treatments, the blood samples were collected by cardiac puncture for estimation of lipid profile.

STATISTICAL ANALYSIS

Data were expressed as Mean ± SEM and statistical analysis was carried out by student’s t test performed using Graph Pad Prism version 5.04.

 

RESULTS:

The study of antidiabetic activity of the stem bark extract of Holoptelea integrifolia was performed in alloxan induced diabetic animals. Originally the study was planned for 21 days, but high mortality rate was seen in alloxan induced diabetic rats and the number of animals surviving 21 days were not sufficient for statistical analysis so that the statistical analysis was done on the data obtained up to day 15.

 

Effect of Holoptelea integrifolia stem bark extract on the blood glucose levels, in alloxan induced diabetic rats.

As result shown in table 1 the effect of stem bark extract of Holoptelea integrifolia on the blood glucose level. It was found that the blood glucose level decrease gradually right through day 1 to day 15 in both extract treated groups i.e. group 3 (250 mg/kg b.w.) and group 4 (500 mg/kg b.w.).  The result was found to be significant after 15 day treatment in group 4 (500 mg/kg b.w.) with respect to the diabetic control group.

 

Effect of Holoptelea integrifolia stem bark extract on the body weight, in alloxan induced diabetic rats.

The effect of stem bark extract of Holoptelea integrifolia on the body weight of alloxan induced diabetic rats was determined shown in table 2. It was found that the loss of body weight in diabetic animals was significantly prevented after 15 day treatment by both test doses i.e. group 3 (250 mg/kg b.w.)) and group 4 (500 mg/kg b.w.) at 1% level level of significance (**P<0.01) and at 5% level of significance (*P<0.05) respectively.

 

Effect of Holoptelea integrifolia stem bark extract on the lipid profile, in alloxan induced diabetic rats.

As per results shown in table 3, the levels of total cholesterol and triglyceride in the animals of group 4 (500 mg/kg b.w.) are lower than those of group 3 (250 mg/kg b.w.) and the levels of HDL are higher than those of group 3 (250 mg/kg b.w.). These values show that the extract is able to correct the higher cholesterol and triglyceride levels characteristic of diabetes mellitus. Similarly it improves the HDL level which decrease in diabetes

 

DISCUSSION AND CONCLUSION:

The present study of antidiabetic activity of stem bark of Holoptelea integrifolia (Roxb.) was taken up on the basis of reports of anti diabetic activity in the leaves of the tree. The protocol consisted of four groups of albino rats (one normal and three diabetic). One of the diabetic group (group 2) served as control and remaining two groups were treated with the bark extract at the dose level of 250 mg/kg orally (group 3) and 500 mg/kg orally (group 4) respectively.

 

The study was originally planned for 21days but the number of animals surviving in 21 days was not sufficient for statistical analysis. The minimum number of animals per group required for applying statistical test could survive only 15 days and therefore statistical conclusions have been drawn on the basis of 15 day data.

 

The results of blood glucose level show that there is a clear trend of reduction in hyperglycemia in the animals treated with the bark extract, right through day 1 to day 15. However this antidiabetic effect was found to be statistically significant only on day 15th in the animals of group 4. Thus the antidiabetic activity of the stem bark extract of Holoptelea integrifolia is apparently statistically significant only after 15 day treatment at 500 mg/kg body weight orally. Although it is not possible to establish the effect on the basis of available data, it is highly probable that group 3 would also show significant antidiabetic activity in the post 15 day period, since the fall in blood glucose level of group 3 animals parallels the response of group 4.

 

 

 

 

Table 1: Effect of Holoptelea integrifolia stem bark extract on the blood glucose levels (mg/dl), in alloxan induced diabetic rats. Values are in mean ± S.E.M.

Groups

Day 0

Day 1

Day 3

Day 7

Day 15

Group 1 (Normal control)

76.33 ± 2.616

76.33 ± 2.231

75.50 ± 1.875

68.00 ± 3.633

69.00 ± 3.582

Group 2 (Diabetic control)

281.7±37.83

242.3 ± 29.69

240.7 ± 30.19

262.5 ± 42.50

286.5 ± 41.50

Group 3 (Test group 250 mg/kg)

240.2 ± 27.00

239.4 ± 27.50

232.8±25.13

200.8 ± 11.54

174.5±9.500

Group 4 (Test group 500 mg/kg)

287.8±46.37

252.8 ± 42.76

245.3 ± 40.24

187.0 ± 9.644

147.5 ± 16.50*

*P<0.05 significant from the diabetic control animal.

 

 

 

 

TABLE 2: Effect of Holoptelea integrifolia stem bark extract on the body weight (mg), in alloxan induced diabetic rats. Values are in mean ± S.E.M

Groups

Day 1

Day 4

Day 7

Day 15

Group 1 (Normal control)

143.0 ± 10.07

145.8 ± 7.683

151.0 ± 7.969

163.8 ± 4.270

Group 2 (Diabetic control)

161.7 ± 9.280

158.3 ± 8.333

143.5 ± 1.500

131.5 ± 1.500

Group 3 (Test group 250 mg/kg)

153.0 ± 10.07

156.3 ± 11.97

149.8 ± 13.29

153.5 ±0.500**

Group 4  (Test group 500 mg/kg)

160.8 ± 5.750

158.3 ± 8.333

153.3 ± 8.333

150.0 ± 5.000*

*P<0.05, **P<0.01 significant from the diabetic control animal

 

 

Table 3: Effect of Holoptelea integrifolia stem bark extract on the lipid profile (mg/dl), in alloxan induced diabetic rats.     

Groups

TC

TG

HDL

VLDL

LDL

Group 1 (Normal control)

67.4

40.8

29.7

13.48

24.22

Group 2 (Diabetic control)

All animals were died

Group 3 (Test group 1 dose 250 mg/kg )

110.3

91.2

15.9

22.06

72.34

Group 4 (Test group 2 dose 500 mg/ kg)

80.15

77.2

18.7

16.03

45.42

 

 

 

 

 

Additional observations on the experimental animals also support beneficial effect of bark extract in diabetic animals. The prevention of loss of body weight in diabetic animals is highly significant on day 15 in the animals of both group 3 and group 4.

 

Test of lipid profile of the animals surviving at the end of study (21 day) could not provide data sufficient to draw statistical conclusion because all animals of diabetic control group (group 2) died before completion of the study. However a comparison of lipid levels in the two test groups and normal control group does reflect the activity of bark extract in correcting the diabetic lipid abnormalities. The levels of total cholesterol and triglyceride in the animals of group 4 are lower than those of group 3 and the levels of HDL are higher than those of group 3. These values show that the extract is able to correct the higher cholesterol and triglyceride levels characteristic of diabetes mellitus. Similarly it improves the HDL level which decrease in diabetes.

 

Overall it may be concluded that Holoptelea integrifolia (Roxb.) bark extract possess the antidiabetic activity which is dose dependent and statistically significant after 15 day treatment at the dose level of 500 mg/kg orally. The extract has also demonstrated the ability to check fall in body weight of diabetic animals. Additionally it appears to correct the lipid abnormalities accompanying diabetes mellitus in a dose dependant manner.

 

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Received on 12.04.2012

Modified on 28.04.2012

Accepted on 07.05.2012                                               

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Research J. Pharmacology and Pharmacodynamics. 4(3): May-June, 2012, 181-184