Hydrogen Sulfide Protects Retinal Pigment Epithelial Cells from Oxidative Stress-Induced Apoptosis and Affects Autophagy

Figure 7

Autophagy is involved in H₂O₂-induced oxidative stress and cell apoptosis. (a) ARPE-19 cells were pretreated with 3-MA for 3 h and then treated with H₂O₂ for 1 h. The protein expression and transform of LC3 I and LC3 II in ARPE-19 cells were analyzed by Western blot. (b–d) The quantitative analyses of LC3 I/GAPDH, LC3 II/GAPDH, and LC3 II/LC3 I are shown. (e) MTT assay was performed to detect the cytotoxicity of different concentrations of the autophagy inhibitor 3-MA for 24 h in ARPE-19 cells. (f) ARPE-19 cells were treated with different concentrations of the autophagy inhibitor 3-MA and 400 μM H₂O₂. MTT assay was performed to examine the viability of ARPE-19 cells after ARPE-19 cells were pretreated with 3-MA for 3 h and then exposed to H₂O₂ for 24 h. (g, i) Cell apoptosis was analyzed with Annexin V-FITC and PI stain by flow cytometry. (h) Cell morphology was examined in a bright field under an inverted fluorescent microscope after ARPE-19 cells were pretreated with 3-MA for 3 h and then exposed to H₂O₂ for 24 h. Scale . Values are the . and versus the control group; ^### versus the H₂O₂ treatment alone group.

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