Protective effects of H2S against H2O2-induced oxidative injury in RGC / 眼科新进展

ABSTRACT

Objective To investigate the protective effects and possible mechanism of hydrogen sulfide (H2S) against oxidative stress injury induced by hydrogen peroxide (H2O2) in retinal ganglion cell-5 (RGC-5).Methods RGC-5 cells were divided into four groupsRGC-5 group (normal control group),RGC-5 + H2O2 (RGC-5 were cultured in 500 μmol · L-1 H2O2 for 24 hours) group,RGC-5 + NaHS (RGC-5 were cultured in 50 μmol · L-1 NaHS for 30 minutes) + H2O2 (RGC-5 were cultured in 500 μmol · L-1 H2 O2 for 24 hours) group,and RGC-5 + NaHS (RGC-5 were cultured in 50 μmol · L-1 NaHS for 30 minutes) group.Western blots were applied to measure the expression of cytochrome c (Cyt.c) and optic atrophy 1 (OPA1).The fluorescent dye JC-1 assay was chosen to detect the mitochondrial membrane potential (△Ψm).Furthermore,transmission electron microscope was used to observe the morphology of mitochondria.Results Compared with RGC-5 group,the expression of Cyt.c in RGC-5 + H2O2 group decreased in mitochondria,and increased in cytoplasm (all P ＜ 0.05),but there was no statistical difference between RGC-5 group and RGC-5 + NarHS + H2O2 group (all P ＞0.05).Compared with RGC-5 group,the expression of Cyt.c in RGC-5 + NaHS group increased in mitochondria,and decreased in cytoplasm (all P ＜ 0.05).Compared with RGC-5 group,the expression of OPA1 in RGC-5 + H2O2 group decreased in mitochondria,and increased in cytoplasm (all P ＜ 0.05).In RGC-5 + NaHs + H2O2 group and RGC-5 + NaHS group,the expression of OPA1 within and outside the mitochondria had no significant difference compared with RGC-5 group (all P ＞ 0.05).Compared with other three groups,the mitochondrial membrane potential in RGC-5 + H2O2 group obviously decreased,but there was no statistical difference among other three groups (P ＞ 0.05).The mitochondria were globular swelled in RGC-5 group,but in other three groups,the mitochondria had slightly swelled.Conclusion H2S can protect the mitochondrial morphology and functions of RGC? 5 from H2O2-induced oxidative stress via preventing OPA1 release from mitochondria.