Shenfu injection protects human ECV304 cells from hydrogen peroxide via its anti-apoptosis way.

Ethnopharmacological relevance The pathogenesis of thromboangiitis obliterans (TAO) has not been fully elucidated until now. Shenfu injection (SFI), a traditional Chinese formula has been widely used clinically for the treatment of cardiovascular diseases for more than two decade. Our previous results first suggested that SFI can cause a significant therapeutic effect on experimental TAO model rats. This experiment was designed to further investigate the protective effect of SFI on VEC damaged by hydrogen peroxide (H 2 O 2 ) oxidative stress in vitro . Meterials and methods The cell viability was evaluated by the MTT assay, the activities of SOD and GSH-PX and the content of MDA in the supernatants of the cultured ECV304 cells were evaluated by a colorimetry method, cell apoptosis was detected by flow cytometry and an AO/EB double staining method. The protein expressions of Bcl2, Bax and caspase-3 were examined by Western blotting. Results When compared with control group, lower survival rate of ECV304 cells was observed in H 2 O 2 group ( p <0.01) ; 20 μl/ml, 30 μl/ml and 40 μl/ml SFI increased the survival rate of ECV304 cells under H 2 O 2 oxidative stress ( p <0.05 and p <0.01). The activities of SOD and GSH-PX were higher and MDA level was lower in H 2 O 2 group than those in control group. These effects of H 2 O 2 on SOD, GSH-PX activities and MDA content were reversed by SFI in concentration-dependent way ( p <0.05 and p <0.01). Flow cytometry and AO–EB double staining discovered that SFI pretreatment inhibited the ECV304 cells apoptosis. The protein expression of caspase3 in 30 μl/ml and 40 μl/ml SFI groups significantly decreased whereas Bcl2 protein expressions in 20 μl/ml, 30 μl/ml and 40 μl/ml SFI groups were higher than H 2 O 2 group, with Bax protein expression much lower than H 2 O 2 group ( p <0.05 and p <0.01). Conclusions Our findings suggest that SFI could prevent the ECV304 cells against H 2 O 2 oxidative-stress by enhancing antioxidant enzyme activities, reducing the membrane lipid peroxidation, as well as upregulating antiapoptotic and downregulating apoptosis protein expressions. [ABSTRACT FROM AUTHOR]