Salvianolate inhibits reactive oxygen species production in H2O2-treated mouse cardiomyocytes in vitro via the TGFβ pathway

Aim: To investigate the effects of salvianolate, a water-soluble active compound from Salvia miltiorrhiza Bunge, on reactive oxygen species (ROS) production in mouse cardiomyocytes in vitro. Methods: Primary ventricular cardiomyocytes were prepared from neonatal mouse. The cell viability was determined using MTT assay. Culture medium for each treatment was collected for measuring the levels of NO, iNOS, total antioxidant capacity (TAOC) and transforming growth factor beta 1 (TGF beta 1). TGF beta 1 and Smad2/3 expression in the cells was detected with Western blotting. Results: H2O2 (1.25 mmol/L) did not significantly affect the cell viability, whereas the high concentration of salvianolate (5 g/L) alone dramatically suppressed the cell viability. Treatment of the cells with H2O2 (1.25 mmol/L) markedly increased ROS and iNOS production, and decreased the levels of NO, TAOC and TGF beta 1 in the culture medium. Furthermore, the H2O2 treatment significantly increased TGF beta 1 and Smad2/3 expression in the cells. Addition of salvianolate (0.05, 0.1, and 0.5 g/L) concentration-dependently reversed the H2O2-induced alterations in the culture medium; addition of salvianolate (0.05 g/L) reversed the H2O2-induced increases of TGF beta 1 and Smad2/3 expression in the cells. Blockage of TGF beta 1 with its antibody (1 mg/L) abolished the above mentioned effects of salvianolate. Conclusion: Salvianolate inhibits ROS and iNOS production and increases TAOC and NO levels in H2O2-treated cardiomyocytes in vitro via downregulation of Smad2/3 and TGF beta 1 expression. High concentration of salvianolate causes cytotoxicity in mouse cardiomyocytes.