Objective To investigate the effect of oxidative stress on the growth and development of human ovarian granu-losa cells.Methods Human ovarian granulosa cells KGN were cultured in vitro and treated with different concentrations of hydrogen peroxide(H2O2),which were measured for the cell proliferation by CCK8 assay,detected for the production of intracellular reactive oxygen species(ROS)and the cell apoptosis by flow cytometry,and determined for the expression of proteins related to autophagy and apoptosis by Western blot.Results With the increasing concentrations of H2O2,the prolife-ration of granulosa cells was significantly inhibited in a dose-dependent manner(F=4.906,4.825,4.653,4.614 and 4.587,respectively,each P＜0.001).The intracellular ROS level increased with the increase of H2O2 concentration,and the apoptosis rate also increased continuously.After the treatment of N-acetyl-L-cysteine(NAC),the ROS level recovered and the apoptosis rate decreased,indicating that cell apoptosis was correlated with ROS release.After the treatment of H2O2,the expression levels of LC3-Ⅱ and cleaved-caspase 3 in KGN cells increased significantly(t=4.809 and 5.789,respectively,each P＜0.05),while the expression levels of LC3-Ⅰ,BCL-2 and P62 decreased significantly(t=4.014,3.982 and 4.415,respectively,each P＜0.05),which indicated the inducing effect of H2O2 on the cell autophagy and apoptosis of KGN cells.Conclusion H2O2 can inhibit the proliferation of human ovarian granulosa cells and induce their cell autophagy and apoptosis.Oxidative stress can inhibit the growth and development of human ovarian granulosa cells,decreased the quality of oocytes and the ovarian reserve function.
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