摘要
目的:探索维生素B12预处理对H2O2诱导下的神经细胞损伤的保护作用及自噬和凋亡蛋白表达的影响。方法:将大鼠肾上腺髓质嗜铬瘤分化细胞(PC12)分为对照组(Control组)RPMI 1640培养、损伤组(H2O2组)RPMI 1640培养并加入200μmol/L的H2O2刺激,维生素B12保护组(H2O2+VitB12组)RPMI 1640培养加入200μmol/L维生素B12预处理和200μmol/L的H2O2刺激。24 h后检测细胞的活性,Western blot和免疫荧光检测凋亡及自噬相关蛋白表达水平。结果:与Control组比较,H2O2组的细胞存活率、细胞DNA复制水平降低,Bcl-2和P62蛋白的表达量下降,Bax、ATG-5和LC3蛋白的表达量明显上升,胞浆内LC3的激活数量上升(均P<0.05)。与H2O2组比较,H2O2+VitB12组细胞存活率和细胞DNA复制水平上升,Bcl-2和P62蛋白的表达量上升,Bax、ATG-5和LC3蛋白的表达量显著下降,胞浆内LC3的激活数量下降(均P<0.05)。结论:维生素B12能通过调控自噬抑制氧化应激诱导下的细胞凋亡。
Objective:To explore the protective effect of Vitamin B12 pretreatment on PC12 cell injury induced by hydrogen peroxide and of its influence on autophagy and apoptotic protein expression.Methods:PC12 cells were divided as the control group(Control)with RPMI 1640 culturethe injury group(H2O2)with RPMI 1640 culture plus 200μmol/L hydrogen peroxide stimulation and the Vitamin B12 protection group(H2O2+Vit B12)with RPMI 1640 culture plus 200μmol/L vitamin B12 and 200μmol/L hydrogen peroxide stimulation.Cell growth was examined 24 h later,and apoptosis and autophagy-related protein expression were detected by Western blot.Results:Compared with the Control group,the cell survival rate,the level of cell DNA replication,the expression level of Bcl-2 and P62 protein,the expression level of Bax,ATG-5 and LC3 protein,and the activation number of LC3 in cytoplasm increased significantly in the injury group(all P<0.05).Compared with the injury group,the cell survival rate and the level of cell DNA replication in the vitamin B12 protected group increased,the expression level of Bcl-2 and P62 protein increased,the expression level of Bax,ATG-5 and LC3 protein significantly decreased,and the activation number of LC3 in the cytoplasm decreased(all P<0.05).Conclusion:Vitamin B12 can inhibit oxidative stress-induced apoptosis by regulating autophagy.
作者
叶璐霞
徐莉菲
薛榆洁
童鹤艳
何逸昉
李校堃
YE Luxia;XU Lifei;XUE Yujie;TONG Heyan;HE Yifang;LI Xiaokun(School of Pharmaceutical Sciences,Wenzhou Medical University,Wenzhou 325035,China)
出处
《温州医科大学学报》
CAS
2020年第6期449-453,共5页
Journal of Wenzhou Medical University
基金
国家级大学生创新创业训练计划立项资助项目(201810343017)。
