摘要
目的:白藜芦醇对缺氧缺糖/复氧复糖(OGD/R)诱导的PC12细胞损伤及凋亡的影响及作用机制。方法:以PC12细胞建立OGD/R自噬性损伤模型,设置空白对照组、模型组、不同浓度(5、10μmol/L)白藜芦醇组,自噬抑制剂(3-MA)组。使用CCK-8试剂盒检测PC12的增殖能力;流式细胞术检测各组细胞凋亡率;ELISA检测各组细胞培养液中TNF-α和IL-6的浓度变化;DHE荧光探针检测ROS的变化;特定试剂盒检测乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)、还原型谷胱甘肽(GSH-PX)含量;Westernblot法检测各组mTOR的磷酸化情况及LC3-Ⅱ、Beclin-1蛋白表达。结果:与空白对照组相比模型组能够显著降低PC12的增殖能力,增加细胞中ROS的含量和凋亡率,诱导TNF-α和IL-6分泌,LDH、MDA含量显著增加,SOD、GSH-PX活性显著下降,而P-mTOR/mTOR、LC3-Ⅱ、Beclin-1表达增加(P<0.01)。与模型组比较,不同浓度白藜芦醇作用PC12时,细胞存活率逐渐上升,ROS含量及凋亡率逐渐下降,LDH、MDA含量减少,SOD、GSH-PX活性随之上升,TNF-α和IL-6分泌显著下降,P-mTOR/mTOR、LC3-Ⅱ、Beclin-1表达进一步增加(P<0.01)。自噬抑制剂(3-MA)组上述指标的检测结果与白藜芦醇组相反。结论:白藜芦醇可能部分通过激活mTOR/自噬通路对神经细胞发挥保护作用。
Objective:To investigate the effect of Resveratrol on injury and apoptosis of PC12 cells induced by OGD/R and explore the possible mechanism.Methods:PC12 cells were cultured in vitro.OGD/R acted on PC12 cells to induce the injury and apoptosis.The cell proliferation ability of each group was detected by CCK-8 assay.The apoptosis rate of cells in each group was detected by flow cytometry.The concentration of nitric oxide (NO ),TNF-α and IL-6 in the culture medium of each group was detected by ELISA.LDH,MDA,SOD and GSH-PX were detected by specific kits.Detection of ROS changes in Cerebral cortical nerve cells by DHE fluorescence probe.Western blot was used to detect the expression of protein (P-mTOR/mTOR,LC3-Ⅱ,Beclin-1 )in each group.Results :Resveratrol significantly enhanced PC12 cells proliferation ability and increased the activities of antioxidant enzymes (snperoxide dismutase,catalase ),scavenged reactive oxygen species (ROS),inhibited TNF-α,IL-6 secretion.Resveratrol also reduced the rate of apoptosis of cells.Resveratrol upregulated protein (P-mTOR/mTOR.LC3-Ⅱ,Beclin-1 )expression.3-MA (inhibitor of autophagy )could prevent this protection.Conclusion:The protective effect of Resveratrol on PC12 cells injury and apoptosis induced by OGD/R via activating mTOR/autophagy.
作者
吴印华
胡万保
WU Yinhua;HU Wanbao(Department of Traditional Chinese Medicine Dongfeng Huaguo Hospital,Hubei Shiyan,442000,China)
出处
《中医药临床杂志》
2019年第2期298-303,共6页
Clinical Journal of Traditional Chinese Medicine
基金
湖北省教育厅科学基金(B2013117)
