Xu Jing, Fan Xiaoyu, Zhu Mou, Zhao Zhenglin, Qi Xiaodan, Gao Han, Liu Ying, Shi Yan, Zhang Chunjing
School of Medical Technology, Qiqihar Medical University, Qiqihar, 161006 China.
Indian J Microbiol. 2023 Jun;63(2):173-180. doi: 10.1007/s12088-023-01065-w. Epub 2023 Jun 2.
To investigate the effect of Ginsenoside Rg3 on insulin secretion in mouse MIN6 cells and the possible mechanism. The cultured mouse pancreatic islet MIN6 cells were divided into control group (NC), Rg3 group (Rg3, 50 μg/L), high glucose group (HG, 33 mmol/L), High glucose and Rg3 group (HG + Rg3), after 48 h of continuous culture, CCK-8 was used to detect cell viability; mouse insulin enzyme-linked immunoassay kit to detect insulin release; ATP content detection kit to detect ATP; DCFH-DA to detect intracellular reactive oxygen species (ROS) levels; total glutathione (T-GSH)/oxidized glutathione (GSSG) assay kit to detect the ratio of GSH/GSSG; Using the mitochondrial membrane channel pore (MPTP) fluorescence detection kit in MIN6 cells and collect the intensity of green fluorescence; Western blot to detect the expression of antioxidant proteins Glutathione reductase (GR). The results showed that compared with the NC group, the cell viability of the HG was decreased ( < 0.05), insulin release decreased ( < 0.001), ATP content decreased significantly ( < 0.001), and ROS content increased ( < 0.01), the GSH/GSSH ratio of pancreatic islet cells decreased ( < 0.05),the green fluorescence intensity decreased ( < 0.001), indicating that the permeability of mitochondria increased and the content of antioxidant protein in the cells decreased ( < 0.05). Compared with the HG group, the cell viability of the HG + Rg3 group was significantly increased ( < 0.05), the amount of insulin released was significantly increased ( < 0.001), ATP content was significantly increased ( < 0.01), and the ROS content was significantly decreased ( < 0.01), GSH/GSSH ratio increased significantly ( < 0.05), the green fluorescence intensity was increased ( < 0.001), indicating that the permeability of mitochondria decreased and antioxidant protein GR content increased significantly ( < 0.05). Taken together, our results suggest that Rg3 has an antioxidant protective effect on mouse pancreatic islet cells damaged by high glucose and maintains pancreatic islet cell function and promotes insulin secretion.
探讨人参皂苷Rg3对小鼠MIN6细胞胰岛素分泌的影响及其可能机制。将培养的小鼠胰岛MIN6细胞分为对照组(NC)、Rg3组(Rg3,50μg/L)、高糖组(HG,33mmol/L)、高糖+Rg3组(HG+Rg3),连续培养48h后,采用CCK-8检测细胞活力;用小鼠胰岛素酶联免疫分析试剂盒检测胰岛素释放量;用ATP含量检测试剂盒检测ATP;用DCFH-DA检测细胞内活性氧(ROS)水平;用总谷胱甘肽(T-GSH)/氧化型谷胱甘肽(GSSG)检测试剂盒检测GSH/GSSG比值;在MIN6细胞中使用线粒体膜通道孔(MPTP)荧光检测试剂盒并收集绿色荧光强度;用蛋白质免疫印迹法检测抗氧化蛋白谷胱甘肽还原酶(GR)的表达。结果显示,与NC组相比,HG组细胞活力降低(<0.05),胰岛素释放量减少(<0.001),ATP含量显著降低(<0.001),ROS含量增加(<0.01),胰岛细胞GSH/GSSH比值降低(<0.05),绿色荧光强度降低(<0.001),表明线粒体通透性增加,细胞内抗氧化蛋白含量降低(<0.05)。与HG组相比,HG+Rg3组细胞活力显著增加(<0.05),胰岛素释放量显著增加(<0.001),ATP含量显著增加(<0.01),ROS含量显著降低(<0.01),GSH/GSSH比值显著增加(<0.05),绿色荧光强度增加(<0.001),表明线粒体通透性降低,抗氧化蛋白GR含量显著增加(<0.05)。综上所述,我们的结果表明Rg3对高糖损伤的小鼠胰岛细胞具有抗氧化保护作用,可维持胰岛细胞功能并促进胰岛素分泌。