过氧化氢诱导人卵巢颗粒细胞氧化应激模型的建立
发布时间：2019-06-17 10:55
【摘要】：目的探讨构建过氧化氢(H_2O_2)诱导人卵巢颗粒细胞氧化应激模型的方法。方法不同浓度H_2O_2处理人卵巢颗粒细胞COV434,采用Western blot检测Caspase-3表达,流式细胞仪和DAPI/TUNEL双染色法检测细胞凋亡情况,H2DCF-DA检测细胞内活性氧族(ROS)水平,通过量效关系和时效关系来确定H_2O_2的最佳作用条件。结果 1~1.5 mmol/L浓度H_2O_2处理COV434细胞2~4h后Caspase-3表达量升高最明显。随着H_2O_2浓度增加及作用时间的延长细胞凋亡率不断升高。细胞内ROS水平随着H_2O_2浓度的增加而增加。1.5mmol/L浓度H_2O_2处理1h后细胞内ROS水平上升最明显。结论 H_2O_2诱导人卵巢颗粒细胞氧化应激模型的最适宜浓度为1~1.5 mmol/L,采用细胞凋亡情况评价作用时间是2~4h,采用ROS评价作用时间为1h。
[Abstract]:Objective to establish a oxidative stress model induced by hydrogen peroxide (H_2O_2) in human ovarian granulosa cells. Methods the expression of Caspase-3 in human ovarian granulosa cells treated with different concentrations of H_2O_2 was detected by Western blot, the apoptosis was detected by flow cytometry and DAPI/TUNEL double staining, the level of intracellular active oxygen group (ROS) was detected by H2DCF-DA, and the optimum action conditions of H_2O_2 were determined by dose-effect relationship and time-effect relationship. Results 1 the expression of Caspase-3 in COV434 cells increased most significantly after treatment with H_2O_2 at the concentration of 1.5 mmol/L for 2 h. The apoptosis rate increased with the increase of H_2O_2 concentration and the prolongation of treatment time. The intracellular ROS level increased with the increase of H_2O_2 concentration. 1.5 mmol/L H_2O_2 treatment for 1 hour increased the intracellular ROS level most significantly. Conclusion the most suitable concentration of H_2O_2 induced oxidative stress model of human ovarian granulosa cells is 1 鈮，
本文编号：2500935
[Abstract]:Objective to establish a oxidative stress model induced by hydrogen peroxide (H_2O_2) in human ovarian granulosa cells. Methods the expression of Caspase-3 in human ovarian granulosa cells treated with different concentrations of H_2O_2 was detected by Western blot, the apoptosis was detected by flow cytometry and DAPI/TUNEL double staining, the level of intracellular active oxygen group (ROS) was detected by H2DCF-DA, and the optimum action conditions of H_2O_2 were determined by dose-effect relationship and time-effect relationship. Results 1 the expression of Caspase-3 in COV434 cells increased most significantly after treatment with H_2O_2 at the concentration of 1.5 mmol/L for 2 h. The apoptosis rate increased with the increase of H_2O_2 concentration and the prolongation of treatment time. The intracellular ROS level increased with the increase of H_2O_2 concentration. 1.5 mmol/L H_2O_2 treatment for 1 hour increased the intracellular ROS level most significantly. Conclusion the most suitable concentration of H_2O_2 induced oxidative stress model of human ovarian granulosa cells is 1 鈮，
本文编号：2500935
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