2Department of Pharmacy, North south University, Dhaka, Bangladesh
3Consultant neurosurgeon, Gouri Devi Hospital and Research Institute Durgapur, MBBS, MS, General surgery, MCH neurosurgery from PGI Chandigarah), India
The leaves of Antidesma bunius linn. Were collected from Lawchapra, Bakshiganj, Jamalpur, Bangladesh in August, 2016, and identified by an expert taxonomist. A voucher specimen was submitted to the national herbarium, Mirpur, Dhaka, Bangladesh. Accession number: DACB 43490.
In cold extraction process powdered plant materials are submerged in a suitable solvent or solvent system in an air tight flat bottom container for seven days, with occasional shaking and stirring. The major portion of plant materials will be dissolved in the solvent. Solvent is then separated from dispersed materials and evaporated to get desired extract. Approximately 400gm of powder were soaked in 1.7 liter of methanol for about 14 days at room temperature with occasional shaking. After 14 days the solution was filtered using cotton filter and Whitman’s filter paper. The filtrates (methanolic extract) obtained were evaporated first using rotary evaporator & then placed in a water- bath until dried or being semisolid. It rendered a gummy or semisolid concentrates and were designated as crude extracts of methanol. Finally, the extract was stored at room temperature.
Alloxan (Hydrate, Lobachemie PVT. LTD.), metformin (COMET tablet, Square Pharmaceuticals Ltd, Bangladesh), Blood samples analyzed for blood glucose content by using Ez Smart 168 (Tyson Bioresearch, Inc. Chu-Nan, Taiwan) glucose test meter. Total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL), high density lipoprotein (HDL) kits were purchased from Human Gesellschaft fur Bio-chemical mbH-Wiesbaden, Germany. Saline water was purchased from local market. Other chemicals were obtained from local sources and were of analytical grade.
Albino rats of either sex, aged 4-5 weeks, obtained from the Animal Resource Branch of the International Centre for Diarrhoeal Diseases and Research, Bangladesh (ICDDR, B) were used for the experiment. They were housed in standard polypropylene cages and kept under controlled room temperature (25 ± 20C; relative humidity 60-70%) in a 12h light-dark cycle were used for the study and animals were fed ICDDR, B formulated rodent food and water. The protocol of the study was approved by the institutional animal ethics committee.
Albino rats were randomly assigned into group I, II, III, IV, V; 4 rats are selected in each group for the respective one week treatment for the determination of blood glucose, lipid profile test studies.
Group II: Diabetic Control; alloxan monohydrate 150 mg/kg b.w. was administered intraperitoneally to rats.
Group III: Diabetic + metformin, 100 mg/kg b.w.
Group IV: Diabetic + Plant Extract, 250 mg/kg b.w.
Group V: Diabetic + Plant Extract, 500 mg/kg b.w.
Metformin was in white crystal form and freshly soluble in water. The solution of metformin was prepared by dissolving with dimethyl sulfoxide (DMSO) and based on literature searches the doses of metformin were selected which were 100 mg/kg b.w. respectively and was administered intra-peritoneal route to rats at 0.5 ml drug.
Firstly, body weight of each rats were measured. Then required amount of alloxan was measured according to the body weight by following the dose of 150 mg of alloxan per 1000 gm of body weight [20,21]. Then calculated amount of alloxan was dissolved in 0.5 ml of sterile normal saline water. Group II-V animals were allowed to fast for 12 hours were rendered diabetic by injection intraperitoneally a freshly prepared solution of alloxan (150mg/kg/b.w.) in saline water after overnight and drink 10% glucose solution to overcome drug induced hypoglycemia. After 24 hours blood glucose content was measured by using glucose monitoring system by the blood sample which was collected from the tail vein of the rats. Animals got the blood glucose levels above 7.0 mmol/L was selected for the study.
The parameters of blood glucose (BG) level, TG, TC, LDL Cholesterol, HDL Cholesterol and body weight were measured after one week of drug treatment to study the effect of methanolic extract of A. bunius leaves (250 & 500 mg/kg BW), Metformin (100mg/kg BW) were applied in alloxan- induced diabetic rats. Metformin was used as standard antidiabetic agent. In order to determine hypoglycemic activity blood samples are collected from tail-veins of rats and tested for blood glucose level by glucometer at 0th, 3rd,5th and 7th day of treatment. Following chart shows the effect of methanolic extracts of the leaves of A. bunius on blood glucose levels of Long Evan rats after one week of continuous treatment. Sequential injection of alloxan caused a significant increase in blood glucose concentration in all group of rats compared with their respective baseline blood glucose and to control values. At all-time points, blood glucose concentration remain unchanged in normal rats treated with distilled water. However, oral administration of the plant extracts (250 & 500mg/kg) as well as metformin (100mg/kg) to diabetic rats decreased in blood glucose concentrations. The blood glucose concentration decrease in case of metformin (66.46%) and A. bunius extract of 250 and 500mg/kg (29.57% and 62.8%) respectively.
|Groups||Fasting Blood glucose level /(mol/l)|
|Days of treatment|
|At the time of grouping||Day 0||Day 3||Day 5||Day 7|
|Control||4.3 ± 0.27||4.6 ± 0.28||3.9 ± 0.20||3.7 ± 0.23||4.0 ± 0.22|
|Untreated diabetic||4.6 ± 0.34||8.2 ± 0.37||9.1 ± 0.32||13.4 ± 0.39||16.4 ± 0.37|
|5.13 ± 0.32||9.0 ± 0.311||6.21 ± 0.37||4.4 ± 0.41||5.5 ± 0.34|
|Diabetic+ Antidesma bunius (250mg/kg)||6.2±0.28||11.4±0.36||11.5±0.32||12.15±0.30||11.55±0.32|
|Diabetic+ Antidesma bunius (500mg/kg)||5.3±0.31||8.3±0.41||8.0±0.22||6.4±0.37||6.2±0.35|
Showed that the effect of the A. bunius extract on TG, TC, HDL, LDL in alloxan induced diabetic rats. After alloxan induced, the result showed that TG, TC, LDL increased while HDL decreased compared to control rats. The administration of A. bunius extracts (250 & 500mg/kg) and metformin (100mg/kg) decreased TG, TC, LDL levels and significantly increased HDL levels.
Figure 3, 4, 5 and 6: Effect of A. bunius extract on TG, TC, LDL and HDL level in alloxan induced diabetic rats. Data as mean± SEM; n=4 in each group.
|Group||Body weight (g)||Organ weight (mg/g)||(mmol/l) Lipid profile|
|Control||75||93||-24||4.73||3.3 ± 0.30||8.2 ± 0.37||3.3 ± 0.32||7.8 ± 0.10|
|Untreated diabetic||97||90||7.21||4.15||4.4 ± 0.31||9.0 ± 0.16||10.4 ± 0.31||5.3 ± 0.30|
|82||94||-14.63||4.49||2.3 ± 0.30||6.2 ± 0.20||2.2 ± 0.20||9.3 ± 0.31|
|Diabetic + Antidesma bunius (250mg/kg)||104||93||10.33||5.02||6.4 ± 0.31||8.0 ± 0.24||5.4 ± 0.41||7.7 ± 0.16|
|Diabetic + Antidesma bunius (500mg/kg)||99||90||9.49||4.21||4.2 ± 0.20||6.5 ± 0.18||3.8 ± 0.06||9.0 ± 0.20|
The authors are grateful to the Department of Pharmacy, Southeast University, Bangladesh, for their support during the research and also thankful to Bangladesh National Herbarium, Dhaka, Bangladesh for their identification.
It is not applicable.
The protocol of the experiment was approved by the animal ethics committee of the Department of Pharmacy, Southeast University, Dhaka, Bangladesh. The animals care and health were maintained according to the guidelines of National Institutes of Health.
We declare that we have no conflict of interest.
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