Tasmina Rahman
Department of Pharmacy, Jahangirnagar University, Savar, Dhaka- 1342, Bangladesh.
Mohammad Salahuddin Bhuiya
Department of Pharmacy, Jahangirnagar University, Savar, Dhaka- 1342, Bangladesh.
Rakib Hasan
Department of Pharmacy, Jahangirnagar University, Savar, Dhaka- 1342, Bangladesh.
M. S. K. Choudhuri
Department of Pharmacy, Jahangirnagar University, Savar, Dhaka- 1342, Bangladesh.
Ashwagandharishta, Serum Cholesterol, HDL, LDL, Triglyceride
Bangladesh Council of Scientific and Industrial Research (BCSIR)
Development of Host and Medicinal Plants
Medicinal Plants
Drugs, chemicals, and reagents: For the toxicological study, Ashwagandharishta was collected from Sri Kundeswari Aushadhalaya Limited, Chittagong. Ketamine injection was obtained from ACI Pharmaceuticals Limited, Bangladesh. All other reagents, assay kits, and chemicals used in this research work were obtained from Human GmbH, Wiesbaden, Germany. Experimental Animals: Albino rats (Rattus novergicus : Sprague-Dawley strain, 48 weeks old, 70-80 g) of both sexes bred and maintained at the animal house of the Department of Pharmacy, Jahangirnagar University, were used in this toxicological experiment. They were kept in a well-ventilated hygienic experimental animal house under constant environmental and adequate nutritional conditions throughout the period of the experiment. They were fed with rat chow which was prepared according to the formula developed by Bangladesh Council of Scientific and Industrial Research (BCSIR). Water was given ad libitum and the animals maintained at 12 hours day and 12 hours night cycle. All experiments on rats were carried out in absolute compliance with the ethical guide for care and use of laboratory animals approved by Biosafety, Biosecurity and Ethical Committee of Faculty of Biological Sciences, Jahangirnagar University. Experimental design- Acute toxicity study: The acute oral toxicity test was performed for Ashwagandharishta (OECD Guideline, 2000). Sixteen male & female rats (70-80 g body weight) were divided into four groups of four animals each. Different doses (1000 mg/Kg, 2000 mg/Kg, 3000 mg/Kg, and 4000 mg/Kg) of experimental drug (ASG) were administered to them. The dose was divided into two fractions and given within 12 hours. Then all the experimental animals were observed for mortality and clinical signs of toxicity at 1, 2, 3, and 4 hours and thereafter once a day for the next three days following ASG administration. Chronic toxicity studies: A total of 40 males and 40 female rats were randomly assigned to the four groups, namely group I (Control: water), group II-Low Dose (0.625 ml/kg BW of ASG), group III-Medium Dose (5.0 ml/kg BW of ASG), and group IV-High Dose (40.0 ml/kg BW of ASG) consisting of 10 males and 10 females in each group. The animals of the control group were administered with distilled water only as per the same volume as the drug-treated group for 51 days. Ayurvedic medicinal preparation was administered to the rats by intra-gastric syringe at a fixed time daily soon after acclimatization. All the experiments were carried out in absolute compliance with the ethical guideline for the care and use of laboratory animals. Blood samples collection and preparation of serum: At the end of the treatment period (51 days), after 18 hours of fasting, blood samples were collected from post vena cava of the rats anesthetizing with Ketamine (500 mg/Kg body, intra peritoneal). Blood samples were transferred into plain sample tubes immediately for serum generation. It was then centrifuged at 4,000 g for 10 minutes using Bench top centrifuge (MSE Minor, England). The supernatant plasma samples were collected using a dry Pasteur pipette and stored in the refrigerator for further analyses. All analyses were completed within 12 hours of sample collection. Biochemical studies: Serum samples were analyzed for Serum Cholesterol, HDL, LDL, and Triglyceride using spectrophotometer (Vitros–250, Johnson & Johnson) Random Access Multibatch Chemistry Analyzer (USA). Statistical analysis: Data were expressed as Mean ± SEM (Standard Error of the Mean). One-way ANOVA followed by Dunnett’s multiple comparisons was performed to analyze this data set. *p <0.05, **p<0.01, and ***p<0.001 was considered statistically significant, highly significant, and very highly significant respectively. Statistical programs used were SPSS (version 16, IBM software Inc, USA).
Jahangirnagar University J. Biol. Sci. 9(1 & 2): 59-68, 2020 (June & December)
Journal