天麻素改善线粒体损伤抑制LPS诱导的PC12细胞凋亡

Effect of Gastrodin on Mitochondrial Damage and Inhibition of LPS⁃Induced Apoptosis of PC12 Cells

目的:以线粒体损伤为切入点探讨天麻素对LPS诱导PC12细胞凋亡的保护作用。方法:以MTT法确立天麻素最佳干预浓度,设立正常对照组、模型组、依达拉奉(0.005 mol/L)阳性对照组及天麻高(0.2 mol/L)、低(0.02 mol/L)浓度组。使用MTT法初步建立PC12细胞损伤模型,确定损伤条件为1μg/mL LPS刺激细胞48 h, DCFH-DA荧光探针及酶标仪检测ROS水平,JC-1检测线粒体膜电位,流式细胞仪检测mPTP开放程度及细胞凋亡,蛋白免疫印迹法检测细胞内Bax/Bcl-2比例。结果:LPS处理后,PC12细胞中ROS水平显著升高(P<0.001),JC-1与mPTP的检测结果均显示线粒体膜电位显著降低(P<0.01),在使用天麻素干预后,PC12细胞ROS表达显著降低,线粒体膜电位显著升高,细胞凋亡率及Bax/Bcl-2水平显著降低(P<0.05~P<0.01)。结论:天麻素可能通过改善PC12细胞中的线粒体损伤,抑制细胞凋亡。

Objective:To investigate the protective effect of gastrodin on LPS induced apoptosis of PC12 cells using mitochondrial injury as entry point.Methods:The optimal intervention concentration of gastrodin was determined by MTT method,and normal control group,model group,Edaravone(0.005 mol/L)positive control group and gastrodin high(0.2 mol/L)and low(0.02 mol/L)concen⁃traiton groups were set up.The damage model of PC12 cells was established by MTT method,and the damage condition was determined as 1μg/mL LPS for 48 hours.ROS level was detected by DCFH⁃DA fluorescent probe and enzyme label instrument,mitochondrial membrane potential was detected by JC⁃1,mPTP openness and apoptosis were detected by flow cytometry,and the ratio of Bax/Bcl⁃2 in the cells was detected by Western Blot.Results:After LPS treatment,ROS levels in PC12 cells were significantly increased(P<0.001),and the results of JC⁃1 and mPTP showed that the mitochondrial membrane potential significantly decreased(P<0.01).After the inter⁃vention of gastrodin,ROS expression in PC12 cells was significantly decreased and mitochondrial membrane potential was sibnificantly increased,the apoptosis rate and Bax/Bcl⁃2 level significantly decreased(P<0.05 toP<0.01).Conclusion:Gastrodin may inhibit apop⁃tosis by ameliorating mitochondrial damage in PC12 cells.