ASSESSMENT OF ANTI - HYPERGLYCEMIC ACTIVITY OF A POLYHERBAL FORMULATION BS019

Sukumar D. and Brahma Srinivasa Rao Desu Department of Pharmacology, Hindu college of Pharmacy, Guntur, Andhra Pradesh, India. ...................................................................................................................... Manuscript Info Abstract ......................... ........................................................................ Manuscript History Received: 14 October 2019 Final Accepted: 16 November 2019 Published: December 2019


Preparation of Polyherbal formulation BS019
The dried flowers of Hibiscus rosa sinensis, leaves of Momordica charantia and roots of senna occidentalis were washed and cleaned separately. The powdered plant materials were used for the preparation of ethanolic extracts. 500 gms of each plant material was weighed and extracted with 95% ethanol by maceration process separately for 4 days and filtered with watchman filter paper. The extracts were concentrated under reduced pressure and stored in vaccum dessicators for complete removal of solvent. Each extract was weighed and percentage yield was calculated 5 .
The polyherbal formulation which contains equal proportions of the ethanolic extract of Hibiscus rosa sinensis (flowers) Momordica charantia (leaves) and senna occidentalis (roots) was called as BS019.

Qualitative Phytochemical Analysis
Phytochemical analysis of Polyherbal formulation BS019 was carried out by using standard procedures to identify the presence of various phytoconstituents 6 .

In-vivo studies: Experimental animals:
Adult wistar albino rats (150-180 g) of either sex were procured from the laboratory animal house, Hindu College of Pharmacy, Guntur, Andhra Pradesh, India and used in the study. The animals were kept under standard environmental conditions of room temperature (22± 2°C), relative humidity (50% ± 5%) and 12 h light and dark cycle. The animals were housed in the colony cages ( three rats per cage) and provided feed (commercial pellets contain a balanced ration obtained from Mahaveera Enterprises, Hyderabad) and water ad libitum.
All the animals were acclimatized to the laboratory environment 5 days prior to experiment. The animals were fasted overnight just prior to the experiment but allowed free access to drinking water. All the experiments were carried out in accordance with the guidelines of Institutional Animal Ethics Committee. The study was conducted after obtaining ethical committee clearance from the Institutional Animal Ethics Committee No : HCOP/IAEC/PR-1 /2019.

Anti-hyperglycemic activity: hypoglycaemic activity in normal rats
The overnight fasted albino wistar rats weighing (150-200 gm ) were divided in to four groups of six animals in each group. All these agents were given by oral route.

Sample collection:
Blood samples were collected from tail puncture of the rats at 0, 30, 60, 120 min respectively after oral administration. The blood glucose level was determined by electronic Glucometer (one touch glucometer-Nipro premier S) 7

Statistical values:
The values were expressed as Mean ± SEM. The data was analyzed by using one way ANOVA followed by Dunnets's test and the values p < 0.05 were considered significant (Table 1). Values are Mean ± SEM (n=6). One way ANOVA followed by Dunnets's *p<0.05 , **p<0.01 , ***p<0.001,when compared to vehicle treated (control) animals.

Alloxan induced diabetic model: Preparation of alloxan solution:
Alloxan was dissolved in saline and a single intra-peritonial injection was administered within five minutes to avoid degradation.

Alloxan induced diabetic model:
Alloxan monohydrate will be first weighed individually for each animal according to its weight and then solubilized in 0.2ml saline just prior to injection. Diabetes was induced by injecting alloxan at a dose of 150mg/kg body weight intra-peritonially. After 1hr of alloxan administration, the animals were given feed and 5% dextrose solution was also given in feeding bottle for a day to overcome the early hypoglycemic phase.

Physical parameters: Determination of body weight:
Body weight of the entire animal in each group was noted on the 0 th and 7 th day of the experimental period. The weight difference was calculated.

Estimation of biochemical parameters:
The blood glucose was determined by electronic Glucometer (one touch glucometer-Nipro premier S). Cholesterol, and Triglycerides were estimated using commercially available kits 8 .

Statistical analysis:
The values were expressed as Mean ± SEM. The data was analyzed by using one way ANOVA followed by Dunnets's test and the values p < 0.05 were considered significant ( Table 2,3,4).

Results & Discussion:-
Traditionally, herbs used for the treatment of disease and disorders. Among other disorders, Diabetic mellitus is a chronic disorder and is a major public heath problem in the developed as well as developing countries caused by partial or complete insulin deficiency, resulting in hyperglycaemia leading to acute and chronic complications . Synthetic drugs produces serious side effects.
Preliminary phytochemical analysis revealed the presence of carbohydrates, alkaloids, tannins, phenolic compounds, volatile oils, flavonoids and glycosides in BS019. The Alloxan induced diabetic rat is one of the animal models of human diabetes mellitus. Diabetes arises from irreversible destruction of pancreatic β cells, causing reduction of insulin secretion. Glibenclamide, a standard hypoglycemic agent was taken for comparison of the glucose lowering effectiveness of the polyherbal formulation BS019. The present study showed that the polyherbal formulation BS019 (at doses 200mg/kg and 400 mg/kg p.o ) had potential anti-hyperglycemic activity when compared to control. At 400 mg/kg BS019 had higher hypoglycemic activity and were similar to the standard drug glibenclamide (10 mg/kg,p.o).The levels of total cholesterol and triglycerides are elevated in diabetes, this is due to uninhibited actions of lipolytic hormones on the fat depots. The oral administration of BS019 also reduced the total cholesterol and triglycerides.
The anti-hyperglycemic activity of BS019 may be due to the presence of alkaloids, tannins, phenolic compounds, volatile oils and flavonoid. However, further studies are necessary to find the exact mechanism of antihyperglycemic activity.

Conclusion:-
From the results of our studies, it can be concluded that Polyherbal formulation BS019 exhibited significant antihyperglycemic activity. The observed effects were nearly equal to the existed familiar standard drug Glibenclamide. However, further studies are necessary to find the exact mechanism of anti-hyperglycemic effect.