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Evaluation of In-vitro Membrane Stabilization and Thrombolytic Activities of Morinda Angustifolia Roxb.

Baharul Islam
Department of Pharmacy, Faculty of Science and Engineering, Southern University Bangladesh, Academic building-III, 739/A, 22-Shahid Mirza Lane (East), Chittagong, Bangladesh.

Afroz Karim
Department of Pharmacy, Faculty of Science and Engineering, Southern University Bangladesh, Academic building-III, 739/A, 22-Shahid Mirza Lane (East), Chittagong, Bangladesh.

Mohammed Mashrur Chowdhury
Department of Pharmacy, Faculty of Science and Engineering, Southern University Bangladesh, Academic building-III, 739/A, 22-Shahid Mirza Lane (East), Chittagong, Bangladesh.

Shatabdi Bhattacharjee
Department of Pharmacy, Faculty of Science and Engineering, Southern University Bangladesh, Academic building-III, 739/A, 22-Shahid Mirza Lane (East), Chittagong, Bangladesh.

Syed Mohammed Tareq
Department of Pharmacy, Faculty of Science and Engineering, Southern University Bangladesh, Academic building-III, 739/A, 22-Shahid Mirza Lane (East), Chittagong, Bangladesh.

Abstract/ Executive Summary:

The aim of the present study was to evaluate the in-vitro membrane stabilization and thrombolytic activities of the methanolic extracts of root and leaves of Morinda angustifolia Roxb. Assessment of in-vitro membrane stabilization activity was carried out by hypotonic solution induced hemolysis method in which both root and leaves extracts along with the standard drug acetyl salicylic acid, significantly (p<0.0001) inhibited hemolysis when compared to control. For instance, root and leave extracts caused maximum inhibition of hemolysis of 40.22% and 21.62% respectively. In in-vitro thrombolytic activity test, 38.26% and 23.66% clots were lysed by root and leaves extract respectively which were also of statistical significance (p<0.0001).

Keywords: Morinda angustifolia Roxb., Membrane stabilization activity, Thrombolytic activity

Location: Forest Research Institute (FRI), Chittagong, Bangladesh.

Start Date:

End Date:

Program Area: Pest Management

Commodity: Medicinal Plants

Objectives:

To evaluate the in-vitro membrane stabilizing and thrombolytic activities of methanolic extracts of root and leaves of M. angustifolia Roxb.

Methodology:

M. angustifolia was collected from Cox’s Bazar Hill Tracts, Bangladesh and was subsequently identified by BangladeshForest Research Institute (BFRI), Chittagong, Bangladesh. To increase the surface area of the plant particles, these were crushed into coarse powders using crushing mill after shade drying for 10 days. Then these coarse powders of root and leaves parts approximately weighing 120 gm, were placed in separate soxhlet extractor (Quickfit, England) for continuous hot extraction with 700 mL methanol for almost 22 hours at 45°C temperature. After extraction, filtration was carried out and the filtrate was made viscous under ceiling fan. Root and leaves parts rendered yield value of 16.45% and 18.73% respectively. All the chemicals used in the tests were of analytical grade and were purchased from Merck, India. The present study was conducted with minor modification of the method. Iso-saline and hypotonic solution were prepared by dissolving 900 mg and 500 mg NaCl in 100 mL water respectively. Particular amount of iso-saline was added to the mixture of fresh human blood and RBC diluting fluid to make 10% RBC suspension. Both the standard drug and the extracts were dissolved in double distilled water. In the different marked test tubes, 1 mL of 10% RBC suspension, 1 mL test substance (e.g. standard drug, extracts) of varying concentrations and 1 mL hypotonic solution were added. In control tubes, in place of test substances, double distilled water was added. Phosphate buffer was used to adjust the pH (7.4±0.2) of the solution mixture. The tubes were placed in a water bath at a temperature of 56°C for 30 min. Then the reaction mixtures were subjected to centrifugation at 2500 rpm for 5 min and after subsequent filtration the absorbance of the supernatant fluids was spectrophotometrically measured at 556 nm. Three times repetition of the test was done. The thrombolytic activity of root and leaves extracts was evaluated following a standard method in which streptokinase (SK) was used as positive control. 5 mL of venous blood was drawn from healthy volunteers, which was distributed in five different previously weighed alpine tubes (0.5 mL/tube) and the tubes were incubated for a period of 45 minutes at 37°C. After 45 minutes when clot formation occurred, the serum portion was removed completely without hampering the clot and each tube was weighed again to determine weight of the clot formed in the alpine tubes (clot weight = weight of clot containing tube – weight of tube alone). The extracts were dissolved in methanol to prepare extract solutions. To the previously marked alpine tubes containing clot of definite weight, 100 µL solutions of root and leaves extracts both at a concentration of 500 µg/100 µL or 5 µg/µL were added separately. 100 µL of streptokinase (SK) and methanol were added as positive and negative control to the marked tubes. All the tubes were then incubated for a period of 90 minutes at 37°C and observed for clot lysis. After incubation, the fluid released was separated and tubes were again weighed to observe the difference in weight after breakdown of clot. The percentage of clot lysis was estimated from the differences in weights measured before and after clot lysis. The results obtained from the tests were expressed as the mean ± SEM and data were analyzed using one-way ANOVA followed by using Dunnett’s t-test. Windows version of SPSS software 20.0 was used to carry out the statistical analysis. A difference was considered significant at p<0.0001.

Source of Information: Der Pharmacia Sinica, 2015, 6(8):40-44, ISSN: 0976-8688

Article Link (Online Journal): www.pelagiaresearchlibrary.com

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

No matter how effective the conventional medications are, they are not completely free of adverse effects and these undesired effects are yet to be minimized along with achieving maximum efficacy. So work on synthesizing more compatible alternatives needs attention; obviously considering patients’ safety. The methanolic extracts of root and leaves of the plant M. angustifolia exhibited some promising activities in the tests. However further researches should be conducted to isolate the active compounds and to carry out subsequent clinical trials.

Knowledge Management: Journal