Research on the mechanism of dioscin-containing Serum against apoptosis of cardiomyocytes induced by hydrogen peroxide / 中国药学杂志

ABSTRACT

OBJECTIVE: To discuss the mechanism of dioscin against apoptosis of cardiomyocytes induced by hydrogen peroxide through Serum Pharmacologic Method. METHODS: The drug-containing serum [0.6 g (crude drug) · kg-1] was prepared by serum pharmacologic method. Cardiomyocytes from neonatal SD rats were cultured in Dulbecco Modified Eagle Medium (DMEM). The primary cultured cardiomyocytes were injured by hydrogen peroxide before giving the drug-containing serum with different concentrations. The cardiomyocyte viability was determined by MTT method. Morphological changes of cardiomyocytes were observed by fluorescence microscope after treating with the drug-containing serum at different concentrations. The anti-apoptotic effect of dioscin was indicated by detecting the activity of caspase-3. The protein expression of Bcl-2 and Bax were semi-quantified by Western-blot method after treating with the drug-containing serum. RESULTS: The cardiomyocyte viability was elevated (P < 0.01) after being treated with different concentrates of drug-containing serum (0.4, 0.8, 1.2 g (crude drug) · kg-1, prepared from the original drug-containing serum). Typical apoptotic features of the cardiomyocytes such as membrane blebbing, cell shrinkage and detachment, and nucleus condensation and fragmentation were dose-dependently improved after being treated with drug-containing serum. The activity of caspase-3 decreased with the increasing concentration of drug. Western blot approach showed that the Bcl-2 level increased meanwhile the Bax decreased. CONCLUSION: Dioscin could increase the viability of the primary cultured cardiomyocytes in hydrogen peroxide induced injury. It could also decrease the activity of caspase-3, improve nucleus condensation and fragmentation, increase Bcl-2 level and decrease the Bax level. Copyright 2012 by the Chinese Pharmaceutical Association.