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Effects of organic extracts and their different fractions of five Bangladeshi plants on in vitro thrombolysis.

Emran TB, Rahman MA, Uddin MM, Rahman MM, Uddin MZ, Dash R, Layzu C - BMC Complement Altern Med (2015)

Bottom Line: Chloroform fractions of T. orientalis, B. monnieri, C. frutescens, B. oleracea and U. sinuata showed highest significant (P < 0.05 and P < 0.001) clot lysis activity viz., 46.44 ± 2.44%, 48.39 ± 10.12%, 36.87 ± .27%, 30.24 ± 0.95% and 47.89 ± 6.83% respectively compared with positive control standard streptokinase (80.77 ± 1.12%) and negative control sterile distilled water (5.69 ± 3.09%).Order of clot lysis activity was found to be: Streptokinase > Chloroform fractions > Methanol (crude) extract > Hydro-methanol fractions > Ethyl acetate fractions > n-hexane fractions > Water.Lower effects of other extracts might suggest the lack of bio-active components and/or insufficient quantities in the extract.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, BGC Trust University Bangladesh, Chittagong, 4000, Bangladesh. talhabmb@gmail.com.

ABSTRACT

Background: The increasingly high incidence of ischemic stroke caused by thrombosis of the arterial vessels is one of the major factors that threaten people's health and lives in the world. The present treatments for thrombosis are still unsatisfactory. Herbal preparations have been used since ancient times for the treatment of several diseases. The aim of this study was to investigate whether herbal preparations possess thrombolytic activity or not.

Methods: An in vitro thrombolytic model was used to check the clot lysis effect of the crude extracts and fractions of five Bangladeshi plant viz., Trema orientalis L., Bacopa monnieri L., Capsicum frutescens L., Brassica oleracea L. and Urena sinuata L. using streptokinase as a positive control and water as a negative control. Briefly, venous blood drawn from twenty healthy volunteers was allowed to form clots which were weighed and treated with the test plant materials to disrupt the clots. Weight of clot after and before treatment provided a percentage of clot lysis.

Results: Using an in vitro thrombolytic model, different fractions of five Bangladeshi medicinal plants namely T. orientalis, B. monnieri, C. frutescens, B. oleracea and U. sinuata showed various range of clot lysis activity. Chloroform fractions of T. orientalis, B. monnieri, C. frutescens, B. oleracea and U. sinuata showed highest significant (P < 0.05 and P < 0.001) clot lysis activity viz., 46.44 ± 2.44%, 48.39 ± 10.12%, 36.87 ± .27%, 30.24 ± 0.95% and 47.89 ± 6.83% respectively compared with positive control standard streptokinase (80.77 ± 1.12%) and negative control sterile distilled water (5.69 ± 3.09%). Other fractions showed moderate to low clot lysis activity. Order of clot lysis activity was found to be: Streptokinase > Chloroform fractions > Methanol (crude) extract > Hydro-methanol fractions > Ethyl acetate fractions > n-hexane fractions > Water.

Conclusions: Our study suggests that thrombolytic activity of T. orientalis, B. monnieri and U. sinuata could be considered as very promising and beneficial for the Bangladeshi traditional medicine. Lower effects of other extracts might suggest the lack of bio-active components and/or insufficient quantities in the extract. In vivo clot dissolving property and active component(s) of T. orientalis and B. monnieri for clot lysis could lead the plants for their therapeutic uses. However, further work will establish whether or not, chloroform soluble phytochemicals from these plants could be incorporated as a thrombolytic agent for the improvement of the patients suffering from atherothrombotic diseases.

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Clot lysis by streptokinase, water, various organic extracts and their different fractions. Effects of drugs on dissolution of clots prepared from blood of normal individuals. Maximum clot lysis (80.77 ± 1.12%) was observed in clot treated with streptokinase (SK). Among herbal drugs chloroform fractions of T. orientalis, B. monnieri, C. frutescens, B. oleracea and U. sinuata showed highest significant (P < 0.05 and P < 0.001) clot lysis activity viz., 46.44 ± 2.44%, 48.39 ± 10.12%, 36.87 ± 1.27%, 30.24 ± 0.95% and 47.89 ± 6.83% respectively. Sterile distilled water (as a negative control) showed 5.69 ± 3.09% clot lysis. Values are mean ± SD, (n = 5); *P < 0.05, **P < 0.001, Dunnet test as compared to control (positive and negative). Data were processed by paired t-test analysis by using SPSS for windows, version 18.0.
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Fig2: Clot lysis by streptokinase, water, various organic extracts and their different fractions. Effects of drugs on dissolution of clots prepared from blood of normal individuals. Maximum clot lysis (80.77 ± 1.12%) was observed in clot treated with streptokinase (SK). Among herbal drugs chloroform fractions of T. orientalis, B. monnieri, C. frutescens, B. oleracea and U. sinuata showed highest significant (P < 0.05 and P < 0.001) clot lysis activity viz., 46.44 ± 2.44%, 48.39 ± 10.12%, 36.87 ± 1.27%, 30.24 ± 0.95% and 47.89 ± 6.83% respectively. Sterile distilled water (as a negative control) showed 5.69 ± 3.09% clot lysis. Values are mean ± SD, (n = 5); *P < 0.05, **P < 0.001, Dunnet test as compared to control (positive and negative). Data were processed by paired t-test analysis by using SPSS for windows, version 18.0.

Mentions: Addition of 100 μL SK (positive control) to the clots along with 90 minutes of incubation at 37°C, showed 80.77 ± 1.12% clot lysis. Sterile distilled water (negative control) treated-clots showed only 5.69 ± 3.09% clot lysis which is very negligible. The mean differences in clot lysis percentage between positive and negative control was very significant (P values < 0.001 and 0.05 respectively). Chloroform fractions of B. monnieri showed the highest (48.39%) significant (P values < 0.001) clot lysis activity among the other extracts. Chloroform fractions of U. sinuata (47.89%) and T. orientalis (46.44%) also gave significant (P values < 0.001) clot lysis which is almost similar to that of B. monnieri chloroform fractions. Chloroform fractions of C. frutiescens and B. oleracea have the moderate but significant (P values < 0.05) clot lysis activity and the values were respectively 36.87% and 30.24%. However, hydro-methanol fractions of T. orientalis, C. frutiescens and B. oleracea have significant clot lysis activity viz. 45.78%, 43.70%, 40.29% respectively (P values < 0.001). n-hexane fractions of only B. monnieri showed moderate also significant (P values < 0.05) clot lysis 32.88% whereas ethyl acetate fractions of T. orientalis (43.29%) and U. sinuata (38.29%) (P values < 0.001 and 0.05 respectively) had significant clot lysis activity. Other ethyl acetate fractions showed very lower clot lyses which were insignificant compared to negative control. Crude methanol extract of all plants except C. frutiescens had significant (P values < 0.001 and 0.05) clot lysis activity. Percent clot lysis obtained after treating the clots with different organic extracts and appropriate controls is shown in Table 2 and represent in Figure 2.Table 2


Effects of organic extracts and their different fractions of five Bangladeshi plants on in vitro thrombolysis.

Emran TB, Rahman MA, Uddin MM, Rahman MM, Uddin MZ, Dash R, Layzu C - BMC Complement Altern Med (2015)

Clot lysis by streptokinase, water, various organic extracts and their different fractions. Effects of drugs on dissolution of clots prepared from blood of normal individuals. Maximum clot lysis (80.77 ± 1.12%) was observed in clot treated with streptokinase (SK). Among herbal drugs chloroform fractions of T. orientalis, B. monnieri, C. frutescens, B. oleracea and U. sinuata showed highest significant (P < 0.05 and P < 0.001) clot lysis activity viz., 46.44 ± 2.44%, 48.39 ± 10.12%, 36.87 ± 1.27%, 30.24 ± 0.95% and 47.89 ± 6.83% respectively. Sterile distilled water (as a negative control) showed 5.69 ± 3.09% clot lysis. Values are mean ± SD, (n = 5); *P < 0.05, **P < 0.001, Dunnet test as compared to control (positive and negative). Data were processed by paired t-test analysis by using SPSS for windows, version 18.0.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4414290&req=5

Fig2: Clot lysis by streptokinase, water, various organic extracts and their different fractions. Effects of drugs on dissolution of clots prepared from blood of normal individuals. Maximum clot lysis (80.77 ± 1.12%) was observed in clot treated with streptokinase (SK). Among herbal drugs chloroform fractions of T. orientalis, B. monnieri, C. frutescens, B. oleracea and U. sinuata showed highest significant (P < 0.05 and P < 0.001) clot lysis activity viz., 46.44 ± 2.44%, 48.39 ± 10.12%, 36.87 ± 1.27%, 30.24 ± 0.95% and 47.89 ± 6.83% respectively. Sterile distilled water (as a negative control) showed 5.69 ± 3.09% clot lysis. Values are mean ± SD, (n = 5); *P < 0.05, **P < 0.001, Dunnet test as compared to control (positive and negative). Data were processed by paired t-test analysis by using SPSS for windows, version 18.0.
Mentions: Addition of 100 μL SK (positive control) to the clots along with 90 minutes of incubation at 37°C, showed 80.77 ± 1.12% clot lysis. Sterile distilled water (negative control) treated-clots showed only 5.69 ± 3.09% clot lysis which is very negligible. The mean differences in clot lysis percentage between positive and negative control was very significant (P values < 0.001 and 0.05 respectively). Chloroform fractions of B. monnieri showed the highest (48.39%) significant (P values < 0.001) clot lysis activity among the other extracts. Chloroform fractions of U. sinuata (47.89%) and T. orientalis (46.44%) also gave significant (P values < 0.001) clot lysis which is almost similar to that of B. monnieri chloroform fractions. Chloroform fractions of C. frutiescens and B. oleracea have the moderate but significant (P values < 0.05) clot lysis activity and the values were respectively 36.87% and 30.24%. However, hydro-methanol fractions of T. orientalis, C. frutiescens and B. oleracea have significant clot lysis activity viz. 45.78%, 43.70%, 40.29% respectively (P values < 0.001). n-hexane fractions of only B. monnieri showed moderate also significant (P values < 0.05) clot lysis 32.88% whereas ethyl acetate fractions of T. orientalis (43.29%) and U. sinuata (38.29%) (P values < 0.001 and 0.05 respectively) had significant clot lysis activity. Other ethyl acetate fractions showed very lower clot lyses which were insignificant compared to negative control. Crude methanol extract of all plants except C. frutiescens had significant (P values < 0.001 and 0.05) clot lysis activity. Percent clot lysis obtained after treating the clots with different organic extracts and appropriate controls is shown in Table 2 and represent in Figure 2.Table 2

Bottom Line: Chloroform fractions of T. orientalis, B. monnieri, C. frutescens, B. oleracea and U. sinuata showed highest significant (P < 0.05 and P < 0.001) clot lysis activity viz., 46.44 ± 2.44%, 48.39 ± 10.12%, 36.87 ± .27%, 30.24 ± 0.95% and 47.89 ± 6.83% respectively compared with positive control standard streptokinase (80.77 ± 1.12%) and negative control sterile distilled water (5.69 ± 3.09%).Order of clot lysis activity was found to be: Streptokinase > Chloroform fractions > Methanol (crude) extract > Hydro-methanol fractions > Ethyl acetate fractions > n-hexane fractions > Water.Lower effects of other extracts might suggest the lack of bio-active components and/or insufficient quantities in the extract.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, BGC Trust University Bangladesh, Chittagong, 4000, Bangladesh. talhabmb@gmail.com.

ABSTRACT

Background: The increasingly high incidence of ischemic stroke caused by thrombosis of the arterial vessels is one of the major factors that threaten people's health and lives in the world. The present treatments for thrombosis are still unsatisfactory. Herbal preparations have been used since ancient times for the treatment of several diseases. The aim of this study was to investigate whether herbal preparations possess thrombolytic activity or not.

Methods: An in vitro thrombolytic model was used to check the clot lysis effect of the crude extracts and fractions of five Bangladeshi plant viz., Trema orientalis L., Bacopa monnieri L., Capsicum frutescens L., Brassica oleracea L. and Urena sinuata L. using streptokinase as a positive control and water as a negative control. Briefly, venous blood drawn from twenty healthy volunteers was allowed to form clots which were weighed and treated with the test plant materials to disrupt the clots. Weight of clot after and before treatment provided a percentage of clot lysis.

Results: Using an in vitro thrombolytic model, different fractions of five Bangladeshi medicinal plants namely T. orientalis, B. monnieri, C. frutescens, B. oleracea and U. sinuata showed various range of clot lysis activity. Chloroform fractions of T. orientalis, B. monnieri, C. frutescens, B. oleracea and U. sinuata showed highest significant (P < 0.05 and P < 0.001) clot lysis activity viz., 46.44 ± 2.44%, 48.39 ± 10.12%, 36.87 ± .27%, 30.24 ± 0.95% and 47.89 ± 6.83% respectively compared with positive control standard streptokinase (80.77 ± 1.12%) and negative control sterile distilled water (5.69 ± 3.09%). Other fractions showed moderate to low clot lysis activity. Order of clot lysis activity was found to be: Streptokinase > Chloroform fractions > Methanol (crude) extract > Hydro-methanol fractions > Ethyl acetate fractions > n-hexane fractions > Water.

Conclusions: Our study suggests that thrombolytic activity of T. orientalis, B. monnieri and U. sinuata could be considered as very promising and beneficial for the Bangladeshi traditional medicine. Lower effects of other extracts might suggest the lack of bio-active components and/or insufficient quantities in the extract. In vivo clot dissolving property and active component(s) of T. orientalis and B. monnieri for clot lysis could lead the plants for their therapeutic uses. However, further work will establish whether or not, chloroform soluble phytochemicals from these plants could be incorporated as a thrombolytic agent for the improvement of the patients suffering from atherothrombotic diseases.

Show MeSH
Related in: MedlinePlus