Toll样受体4介导Aβ_(25-35)诱导的PC12细胞凋亡和线粒体自噬  

Toll-like receptor 4 mediates the apoptosis and mitochondrial autophagy of PC12 cells induced by Aβ_(25-35)

作　　者：张誓伟[1] 赵杰菲 贺显君[2] 乔莹[3] 睢志向 宋彦[1] Zhang Shiwei;Zhao Jiefei;He Xianjun;Qiao Ying;Sui Zhixiang;Song Yan(Department of Psychiatry,Nanyang Second Genera Hospital,Nanyang,473000,China;Department of Psychiatry,ShangqiuFirst People’s Hospital,Shangqiu 476000,China;School of Foreign Languages,Nanyang Institute of Technology,Nanyang 473000,China;Department of Psychiatry,The People’s Hospital of Ningling,Shangqiu 476000,China)

机构地区：[1]南阳市第二人民医院神经内科,南阳473000 [2]商丘市第一人民医院神经内科,商丘476000 [3]南阳理工学院外国语学院,南阳473000 [4]河南省宁陵县人民医院神经内科,商丘476700

出　　处：《中国组织化学与细胞化学杂志》2021年第5期455-461,共7页Chinese Journal of Histochemistry and Cytochemistry

基　　金：河南省医学科技攻关(联合共建)项目(LHGJ20191467)。

摘　　要：目的探讨沉默Toll样受体4(Toll-like receptor 4,TLR4)表达对Aβ诱导的PC12细胞凋亡和线粒体自噬的影响。方法采用Aβ_(25-35)体外培养PC12细胞构建AD细胞模型,并采用TLR4 shRNA慢病毒感染AD细胞。实时荧光定量PCR(qRT-PCR)检测细胞中TLR4 mRNA的表达,CCK-8法、流式细胞仪及荧光分光光度计检测抑制TLR4的表达对AD细胞的存活率、凋亡率及线粒体膜电位的影响。蛋白免疫印迹(Western blot)检测TLR4及自噬相关蛋白LC3II/I、PINK1、parkin的表达变化。结果TLR4在AD细胞中高表达,沉默TLR4能够抑制AD细胞中TLR4的表达。与正常PC12相比,AD细胞的存活率和线粒体膜电位显著降低,而细胞凋亡率及自噬相关蛋白LC3II/I、PINK1和parkin的表达量均显著升高;沉默TLR4能够促进AD细胞的存活及线粒体膜电位的升高,抑制AD细胞的凋亡及自噬相关蛋白LC3Ⅱ/Ⅰ、PINK1和Parkin的表达。结论TLR4在AD细胞中高表达,可能通过抑制AD细胞的线粒体膜电位促进细胞凋亡和线粒体自噬。Objective To investigate the effect of silenced Toll-like receptor 4(TLR4)expression on apoptosis and mitochondrial autophagy in PC12 cells induced by Aβ.Methods PC12 cells were cultured with Aβ_(25-35) to construct AD cell model in vitro.TLR4 shRNA lentivirus was transfected into AD cells to inhibit the expression of TLR4.The expression of TLR4 mRNA was detected by qRT-PCR.The CCK-8 assay,flow cytometry and spectrophotometry were used to detect the effect of TLR4 inhibition on the cell viability,cell apoptosis and mitochondrial membrane potential.The expression of TLR4,and autophagy-related proteins LC3II/I,PINK1,parkin in each group were detected by Western blot.Results TLR4 was highly expressed in the AD cells,and the expression of TLR4 was inhibited by TLR4 shRNA.Compared with PC12 cells,the cell viability and mitochondrial membrane potential were significantly decreased,while the apoptosis rate and the expression of LC3II/I,PINK1 and Parkin were significantly increased in the AD cells.Silencing TLR4 could promote the cell viability and the increase of mitochondrial membrane potential,while inhibit the apoptosis and the expression of LC3Ⅱ/Ⅰ,PINK1 and parkin.Conclusion TLR4 is highly expressed in AD cells,which could promote apoptosis and mitochondrial autophagy by inhibiting mitochondrial membrane potential of AD cells.

关 键 词：TOLL样受体4 阿尔茨海默病 线粒体膜电位 凋亡 自噬 

分 类 号：R749[医药卫生—神经病学与精神病学]