Mohammed Aktar Sayeed
Department of pharmacy, International Islamic University Chittagong, Chittagong-4203, Bangladesh
Mohammad Mamun Ur Rashid*
Department of Pharmacy, Faculty of Life Science, North South University, Dhaka, Bangladesh.
Mohammad Fazlul Kabir
Department of Pharmacy, Faculty of Science and Engineering, International Islamic University Chittagong, Chittagong-4203, Bangladesh
Rashedul Alam
Department of Pharmacy, Faculty of Science and Engineering, International Islamic University Chittagong, Chittagong-4203, Bangladesh
Md. Saiful Islam
Department of Pharmacy, Faculty of Science and Engineering, International Islamic University Chittagong, Chittagong-4203, Bangladesh
Rana Dhar
Department of Pharmacy, Faculty of Science and Engineering, International Islamic University Chittagong, Chittagong-4203, Bangladesh
A. T. M Yusuf
Department of Pharmacy, Faculty of Life Science, North South University, Dhaka, Bangladesh.
Protium serratum, Anti-arthric, Thrombolytic, % lysis of clot, Protein denaturation.
Department of pharmacy, International Islamic University Chittagong, Chittagong-4203, Bangladesh
Development of Host and Medicinal Plants
Medicinal Plants
Plant The leaves of P. serratum were collected from Bandarban district of Chittagong hill tracts region of Bangladesh in 2013 and authenticated by Dr. Shaikh Bokhtear Uddin, Associate Professor, Department of Botany, University of Chittagong, Chittagong-4331, Bangladesh.
Extracts preparation The collected plant was washed thoroughly with water and air dried for a week at 35 to 40°C and pulverized in electric grinder. The obtained powder was successively extracted in methanol (55 to 60°C). The extracts were made to dry by using rotary evaporator under reduced pressure.
In vitro anti-arthritic activity For the evaluation in vitro anti-arthritic activity of P. serratum, the method used was “inhibition of protein denaturation” using diclofenac sodium a standard. The test solution (0.5 ml) consists of 0.45 ml of bovine serum albumin (5% w/v aqueous solution) and 0.05 ml of test solution (methanolic extract of P. serratum). The test control solution (0.5 ml) consists of 0.45 ml of bovine serum albumin (5% w/v aqueous solution) and 0.05 ml of distilled water. Product control (0.5 ml) consists of 0.45 ml of distilled water and 0.05 ml of test solution. Standard solution (0.5 ml) consists of 0.45 ml of bovine serum albumin (5% w/v aqueous solution) and 0.05 ml of diclofenac sodium. Various concentrations (62.5, 125, 250, 500, 1000 μg/ml) of methanolic extact of P. serratum (MEPS) and diclofenac sodium (standard) were taken, respectively. All the solutions were adjusted to pH 6.3 using 1 N HCl. The samples were incubated at 37°C for 20 min and the temperature was increased to keep the samples at 57°C for 3 min. After cooling, 2.5 ml of phosphate buffer was add to the previous solutions. The absorbance was measured using UV-Visible spectrophotometer at 416 nm. The control represents 100% protein denaturation. The results were compared with diclofenac sodium. The percentage inhibition of protein denaturation of different concentrations is tabulated in Table 1. The percentage inhibition of protein denaturation can be calculated as:
% of Inhibition = [100 - (OD of test solution - OD of product control)] × 100
Where OD = optical density
The control represents 100% protein denaturation. The results were compared with diclofenac sodium.
In-vitro thrombolytic test The thrombolytic activity of this extractive was evaluated by the in vitro thrombolytic test (Prasad et al., 2006) using streptokinase as standard. The dry crude extract (10 mg) was suspended in 10 ml of distilled water and it was kept overnight. Then the soluble supernatant was decanted and filtered. Aliquots (5 ml) of venous blood were drawn from healthy volunteers which were distributed in five different pre weighed sterile micro centrifuge tube (1 ml/tube) and incubated at 37°C for 45 min. After clot formation, the serum was completely removed without disturbing the clot and each tube having clot was again weighed to determine the clot weight (Clot weight = weight of clot containing tube – weight of tube alone). To each micro-centrifuge tube containing pre-weighed clot, 100 μl aqueous solutions of different partitionates along with the crude extract was added separately. As a positive control, 100 μl of streptokinase (SK) and as a negative non thrombolytic control, 100 μl of distilled water were separately added to the control tubes. All the tubes were then incubated at 37°C for 90 min and observed for clot lysis. After incubation, the released of fluid was removed and tubes were again weighed to observe the difference in weight after clot disruption. The differences in weights taken before and after clot lysis were expressed as percentage of clot lysis as shown. % of clot lysis = (weight of released clot / clot weight) × 100.
Statistical analysis The significance between % clot lysis by herbal extract by means of weight difference was tested by the paired t-test analysis. Data are expressed as mean ± standard deviation.
Journal of Medicinal Plant Research Vol. 8(16), pp. 615-618, 25 April, 2014
Journal