A Novel H2S Donor 8L Protects H9c2 Cardiomyocytes Against H_2O_2-induced Damage in An NF-E2-related Factor 2-dependent Manner

Aim To explore the effects of a novel H2 S donor( 8L) on oxidative stress-induced damage and the mechanisms underlying NF-E2-related factor 2( Nrf2) in H9c2 cardiomyocytes. Methods H9c2 cardiomyocytes were treated with exogenous reactive oxygen species,hydrogen peroxide( H2O2),to set up an oxidative injury to mimic the in vitro status induced by acute myocardial ischemia-reperfusion. Prior to the treatment with H2O2,the cells were treated with 8L and then its protective action was investigated. In order to test the roles of Nrf2,its selective inhibitor brusatol( BR) was used before H2O2 or 8L. Cell counting kit-8 was used to measure cell viability. Lactate dehydrogenase( LDH) release was assessed by a commercial kit,mitochondrial membrane potential( MMP) was observed by rhodamine123 staining followed by photofluorography and nuclear Nrf2 expression was examined by Western Blot assay. Results Exposure of H9c2 cardiomyocytes to H2O2 ranging from 0 to 600 μmol / L decreased cell viability in a concentration-dependent manner with a lethal concentration of 400 μmol / L. Treatment of the cells with 400 μmol / L H2O2 for 6 h increased LDH release and nuclear Nrf2 expression,while decreased MMP( P 0. 01). Before the treatment with 400 μmol / L H2O2,the cells were preconditioned with 8L at 50,100 and 200 μmol/L for 1 h and the cell viability were enhanced to( 72. 5 ± 6. 3) %,( 83. 1 ± 5. 2) % and( 85. 7 ± 4. 9) % from( 52. 6 ± 4. 3) %,respectively. The pretreatment with 200μmol / L 8L for 1 h significantly attenuated H2O2-induced LDH release( P 0. 01) and MMP loss( P 0. 05),while facilitated H2O2-induced upregulation of nuclear Nrf2 expression( P 0. 01). Pretreatment with 10 μmol / L BR for 1 h not only aggravated H2O2-induced cell injury( P 0. 01) but also partially blocked 8L-induced cell protection( P 0. 01).Conclusion The novel H2 S donor 8L mitigates oxidative stress-induced injury and the mechanisms may be associated with the activation of Nrf2.