表没食子儿茶素没食子酸酯对IL-1β诱导MIN6细胞凋亡的保护作用

Effects of Epigallocatechin gallate on IL-1βinduced MIN6 cells apoptosis

目的探讨表没食子儿茶素没食子酸酯(EGCG)对胰岛细胞的保护作用及机制。方法实验分为正常对照组,IL-1β组,IL-1β+EGCG1组(低浓度),IL-1β+EGCG2组(高浓度)。CCK8检测细胞活性,酶联免疫吸附实验(ELISA)方法检测MIN6细胞基础胰岛素和葡萄糖刺激的胰岛素分泌量,Hochest染色及流式细胞术检测细胞凋亡,JC-1流式细胞术检测线粒体膜电位,比色法测定细胞腺嘌呤核苷三磷酸(ATP)含量,化学荧光法检测细胞活性氧簇(ROS)活性。结果与正常对照组相比,IL-1β组细胞活性降低,基础和葡糖糖刺激胰岛素分泌量显著减少,细胞凋亡明显增加,同时测得细胞线粒体膜电位降低,ATP含量减少,提示细胞线粒体功能损伤,并且ROS活性增加。给予低浓度和高浓度EGCG作用后,与IL-1β组相比,细胞活性明显提高,基础和葡糖糖刺激胰岛素分泌量增加,细胞凋亡率显著减少,线粒体膜电位增加,ATP含量增加,同时ROS活性降低。且IL-1β+EGCG2组作用更强。结论 EGCC可减轻IL-1β诱导的MIN6细胞细胞凋亡率,其机制可能与提高细胞ATP的含量,线粒体膜电位及降低ROS活性有关。

Objective To investigate the effects of Epigallocatechin gallate（EGCG）on IL-1βinduced MIN6 cells apoptosis.M.Methods The experiment group was divided into control group,IL-1βgroup,IL-1β＋EGCG low concentration group and IL-1β＋EGCG high concentration group.Cell activity was detected by CCK8.Insulin secretion was detected by ELISA.cell apoptosis was detected by flow cytometry.The mitochondrial membrane potential was detected by flow cytometry.ATP content and cell activity of ROS were detected by colorimetry and chemiluminescence method.Results Compared with normal group,IL-1βgroup showed much lower cell activity,insulin secretion,cell mitochondrial membrane potential and ATP content,and at the same time IL-1βgroup had significantly higher cell apoptosis and ROS activities.After given EGCG,both low concentration group and high concentration group had higher cell activity,insulin secretion,cell mitochondrial membrane potential and ATP content,at the same time lower cell apoptosis and ROS activities was showed.And the IL-1β＋EGCG high concentration group worked more powerful.Conclusion EGCG has protective effects on IL-1βinduced MIN6 cells apoptosis.Its mechanism may be related to increasing the content of the ATP and mitochondrial membrane potential and protecting mitochondrial function as well reducing the activity of ROS.