稳定转染hSOD1G93A增加NSC34细胞凋亡  

Stably transfection of hSOD1G93A increased apoptosis in NSC34 cells

作　　者：齐伟静[1] 刘亚玲[3] 周晓萌 杨帅[2] 耿丽颖 张欣[1] Qi Weijing;Liu Yaling;Zhou Xiaomeng;Yang Shuai;Geng Liying;Zhang Xin(Department of Neurology,the Baoding No.1 Central Hospital,Hebei 07W00,China)

机构地区：[1]保定市第一中心医院神经内科,河北071000 [2]保定市第一中心医院骨科,河北071000 [3]河北医科大学第二医院神经内科,河北省神经病学重点实验室

出　　处：《脑与神经疾病杂志》2021年第9期544-547,共4页Journal of Brain and Nervous Diseases

基　　金：河北省卫生健康委医学科学研究课题计划(20200265)。

摘　　要：目的探讨稳定转染hSOD1G93A对NSC34细胞凋亡的影响。方法常规体外培养稳定转染空质粒、hSOD1WT或hSOD1G93A质粒的NSC34细胞系,分为3组:空转组、野生组和突变组。应用CCK-8试剂盒和LDH试剂盒来测定各组细胞活力,应用透射电镜观察细胞线粒体形态,通过TUNEL染色来检测细胞凋亡情况,应用Western blotting检测各组细胞凋亡相关蛋白的表达水平。结果与空转组和野生组细胞相比,突变组细胞线粒体形态异常,细胞活力明显下降,凋亡细胞数量增多,差异有统计学意义(P<0.05);且突变组细胞Bcl-2的蛋白表达水平显著下降,Bax的蛋白表达水平均明显增高,差异有统计学意义(P<0.05)。结论稳定转染hSOD1G93A可导致NSC34细胞线粒体形态异常,降低抗凋亡蛋白的表达水平,增加促凋亡蛋白的表达水平,最终导致凋亡细胞增加。Objective To investigate the effect of transfection of hSOD1 G93 A on apoptosis in NSC34 cells.Methods The NSC34 cell lines from three groups(stably transfected with empty,hSOD1 WT or hSOD1 G93 A plasmids)were cultured at routine ways.Cell viability was determine by CCK-8 and LDH,mitochondrial morphology was observed by transmission electron microscopy,cell apoptosis was detected by TUNEL staining,and the protein expression levels of apoptosis-related proteins were detected by western blotting.Results Compared with those in the other two cell lines,the mutant group showed abnormal mitochondrial morphology,the cell viability decreased significantly,and the number of apoptotic cells increased(P<0.05);moreover,the protein expression level of Bcl-2 in the mutant group decreased significantly,and the protein expression levels of Bax increased significantly(P<0.05).Conclusions Stably transfection of hSOD1 G93 A led to mitochondrial structure damage in NSC34 cells,decreased the expression level of anti-apoptotic protein and increased the expression level of pro-apoptotic protein,eventually increased apoptosis in NSC34 cells.

关 键 词：hSOD1G93A NSC34细胞 肌萎缩侧索硬化 凋亡 

分 类 号：R744.8[医药卫生—神经病学与精神病学]