人参皂苷Rg3对H2O2诱导人卵巢颗粒细胞氧化应激损伤的保护作用#br#

摘要：目的　探讨人参皂苷Rg3对过氧化氢（H2O2）诱导人卵巢颗粒细胞（KGN）损伤的保护作用及其可能的作用机制。方法　体外培养KGN，采用不同剂量H2O2孵育细胞2 h，建立H2O2刺激的KGN损伤模型。人参皂苷Rg3预处理24 h，H2O2刺激KGN后，CCK-8检测细胞活力，筛选有效的干预剂量，随后将KGN分为空白组、H2O2模型组、H2O2+Rg3组和Rg3组，采用DCFH-DA荧光探针检测细胞内活性氧（ROS）水平；AnnexinⅤ/PI双染，流式细胞术检测人参皂苷Rg3对H2O2诱导KGN凋亡的影响；Western blot检测凋亡相关蛋白Bcl-2、BAX及细胞色素C的表达。结果　与空白组相比，H2O2模型组细胞活力下降，凋亡比例升高（P＜0.01），ROS水平增高，细胞色素C水平显著升高，Bcl-2/BAX降低（均P＜0.05）。与模型组相比，H2O2+Rg3组能够明显提高细胞活力，减少ROS的产生，降低凋亡细胞比例，降低细胞色素C水平，提高Bcl-2/BAX（均P＜0.05）。与空白组相比，Rg3组无明显差异。结论　人参皂苷Rg3预处理对H2O2诱导的KGN氧化损伤具有保护作用，其机制与减缓ROS过量累积以及抑制线粒体凋亡途径有关。

The protective effect of ginsenoside Rg3 on oxidative stress injuryinduced by H2O2 in KGN cells#br#

Abstract: Objective　To investigate the protective effect of ginsenoside Rg3 on hydrogen peroxide (H2O2)-induced injury of human ovarian granulosa cells KGN, and to explore its possible mechanism. Methods　KGN cells were cultured and then stimulated with different concentrations of H2O2 for 2 hours to establish a model of oxidative damage in KGN cells. Ginsenoside Rg3 pretreated for 24 h followed by stimulation with H2O2. CCK-8 kit was used to detect the cell viability and to screen the effective concentration. KGN cells were divided into 4 groups, control group, H2O2 model group, H2O2+Rg3 group and Rg3 group. The level of intracellular reactive oxygen species (ROS) was detected by DCFH-DA fluorescent probe. AnnexinⅤ/PI double staining and flow cytometry were used to detect the effect of ginsenoside Rg3 on the apoptosis of KGN cells induced by H2O2. Western blot assay was used to detect apoptosis-related proteins BAX and Bcl-2 and the expression of cytochrome C. Results　Compared with the control group, the cell viability decreased in the H2O2 model group, the ratioof apoptosis increased (P＜0.01), the level of ROS increased, the level of cytochrome C increased significantly, and the ratio of Bcl-2/BAX decreased (P＜0.05). Compared with the model group, ginsenoside Rg3 pretreatment for 24 h can significantly increase the cell viability, reduce excessive ROS production, reduce the proportion of apoptotic cells, reduce the level of cytochrome C and increase the expression of Bcl-2/BAX (P＜0.05). There were no significant differences in the above indicators between the control group and Rg3 group. Conclusion Ginsenoside Rg3 preconditioning has a significant protective effect on H2O2-induced oxidative damage in KGN cells, which is related to slowing down ROS excessive accumulation and inhibiting mitochondrial apoptosis pathway.