Hydrogen peroxide-induced oxidative stress model of human ovarian granulose cells
LIU Jian
NIE Guang-ning
YANG Hong-yan
Abstract：Objective To establish the H2O2-induced oxidative stress model in human ovarian granulose cells.Methods Human ovaria granulosa cells (COV434) were treated with H2 O2 at different concentrations.Caspase-3 expression was detected by Western blot.Cell apoptosis was measured by flow cytometry analysis and DAPI/TUNEL double staining method.ROS level in ovaria granulosa cells were measured by flow cytometry after H2DCF-DA probe was loaded.The optimal action condition of H2O2 was confirmed by dose-effect and time-effect relationship.Results Caspase-3 expression increased most significantly after COV434 cells were treated with H2O2 at 1-1.5 mmol/L concentrations for 2-4 h.Following the increasing H2O2 concentration and the lengthening of the action time,cell apoptosis proportion increased.Intracellular ROS level increased with the increasing concentration.ROS level increased most significantly after cells were treated with H2 O2 at 1.5 mmol/L concentrations for 2 h.Conclusion The most appropriate concentration of H2O2 which induced oxidative stress model of human ovarian granulosa cells is 1-1.5 mmol/L,and the action time is 2-4h,while the action time is 1 h when estimating ROS level.
Keywords：hydrogen peroxidehuman ovaria granulosa cells COV434oxidative stress model
Publication Date：2017-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 986-989 )