Proteome Analysis Of The Anti-oxidant Protective Effect Of Salvianolic Acid B's On Endothelial Cells

Salvianolic acid B (Sal B), an antioxidant from traditional Chinese herb Salvia miltiorrhiza Bunge, has been found to exhibit multiple protective effects on vascular cells. In the present study, we showed that Salvianolic acid B (Sal B) was able to protect EA. Hy926 endothelial cells from oxidative damage. To further elucidate the mechanism of the antioxidant protective effect of Sal B on EA. Hy926 endothelial cells, we performed a quantitative proteomics study employing the 16O/18O co-digestion labeling strategy. Protein expression levels were compared in normally cultured endothelial cells, cells pretreated with Sal B and subsequently induced by H2O2 as well as cells only induced by H₂O₂.Among the 125 identified soluble proteins, 9 proteins were considerably differentially expressed in the three experimental groups, including transcriptional and translational regulators, apoptosis inhibitors and anti-oxidant depense proteins. Two transcriptional and translational regulators, ATP-dependent RNA helicase eIF4A-1(eIF4A-I) and Heterogeneous nuclear ribonucleoproteins A2/B1 (hnRNP A2/ hnRNP B1), were up-regulated under oxidative stress in the condition of Sal B pretreatment, compared to the normally cultured cells. As to the apoptosis regulators, the inhibitor of Rho apoptotic pathway—Rho GDP-dissociation inhibitor 1 was up regulated under oxidative stress in the Sal B pretreatment dependent way; and the inhibitor of Caspase3 and Caspase9, High mobility group protein B1 (HMGB1), was up regulated under oxidative stress in the Sal B pretreatment independent way but can be further enhanced by Sal B pretreatment. As to the cellular defense proteins, Heat-shock protein beta-1 (HspB1) and Hsc70-interacting protein (Hip) were up regulated under oxidative stress independent of Sal B pretreatment but further enhanced by Sal B pretreatment; and the cell homeostasis maintaining protein, Ubiquitin carboxyl-terminal hydrolase isozyme L1 (UCH-L1), was up regulated under oxidative stress depend on Sal B pretreatment.In conclusion, Sal B can protect EA. Hy926 cells from oxidative damage through inhibiting the apoptotic signal transduction and the activities of the apoptosis executing molecules, as well as up regulating the expression levels of the cellular defense proteins, all of which were benefited by the up regulations of the transcriptional and translational regulators in the Sal B pretreatment dependent way.