内质网应激在乙型脑炎病毒诱导神经元细胞凋亡中的作用及机制研究  

Role and Mechanism of Endoplasmic Reticulum Stress in Japanese Encephalitis Virus Induced Neuronal Apoptosis

作　　者：王贵霞[1] 曾跃[1] 余婷[1] 林家福[1] 王丽恒[1] 牛绍伟 任亮[1] WANG Guixia;ZENG Yue;YU Ting;LIN Jiafu;WANG Liheng;NIU Shaowei;REN Liang(Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,China)

机构地区：[1]川北医学院附属医院,南充637000

出　　处：《病毒学报》2021年第6期1420-1427,共8页Chinese Journal of Virology

基　　金：南充市科技项目(项目号:NSMC20170415),题目:MBP、MCP-1/FKN在评判乙脑患者及乙脑病毒感染SPF小鼠模型中脑损伤的意义。

摘　　要：内质网应激(Endoplasmic reticulum stress,ERS)的激活与创伤、缺血再灌注等病理刺激引起的神经元凋亡有关,流行性乙型脑炎病毒(JEV)感染能够促进神经元凋亡、激活ERS,但ERS在JEV诱导神经元细胞凋亡中的作用尚不清楚。为了研究ERS在JEV诱导神经元细胞凋亡中的作用及机制,本研究以神经细胞株SH-SY5Y为对象,感染JEV并加用ERS激动剂、ERS抑制剂或转染阴性对照(NC)siRNA、蛋白激酶R样内质网激酶(PERK)siRNA,检测细胞存活率、凋亡率、ERS蛋白PERK、肌醇必需酶-1α(IRE1α)、活化转录因子6(ATF6)及凋亡蛋白C/EBP同源蛋白(CHOP)、含半胱氨酸的天冬氨酸蛋白水解酶12(Caspase-12)、Bcl-2相关X蛋白(Bax)的表达。结果显示:JEV组SH-SY5Y细胞的凋亡率及PERK、CHOP、Caspase-12、Bax的表达水平高于对照组,存活率低于对照组(P<0.05),IRE1α、ATF6的表达水平与对照组比较无显著差异(P>0.05)。与JEV组比较,激动剂组SH-SY5Y细胞的凋亡率及PERK、CHOP、Caspase-12、Bax的表达水平显著增加,存活率显著降低(P<0.05);抑制剂组SH-SY5Y细胞的凋亡率及PERK、CHOP、Caspase-12、Bax的表达水平显著降低,存活率显著增加(P<0.05);与si-NC+JEV组比较,si-PERK+JEV组SH-SY5Y细胞的凋亡率及PERK、CHOP、Caspase-12、Bax的表达水平P显著降低,存活率显著增加(P<0.05)。以上结果表明ERS的PERK通路激活与JEV诱导神经元凋亡有关。Activation of endoplasmic reticulum stress(ERS)relates to neuronal apoptosis induced by trauma,ischemia - reperfusion and other pathological stimuli. Japanese encephalitis virus(JEV)infection can promote neuronal apoptosis and activate ERS,but the role of ERS in JEV induced neuronal apoptosis is still unclear. In order to study the role and mechanism of ERS in JEV induced neuronal apoptosis,SH-SY5 Y was infected with JEV and treated with ERS agonist,ERS antagonist,or transfected with negative control(NC) siRNA or PERK siRNA. The cell survival rate,apoptosis rate,expression of ERS protein PERK,IRE1α,ATF6 and apoptosis protein CHOP,caspase - 12 and Bax were detected. Results showed that the apoptosis rate and the expression levels of PERK,CHOP,caspase - 12 and Bax in SH - SY5 Y cells of JEV group were higher than those of control group,and the survival rate was lower than control group(P<0.05),the expression levels of IRE1 α and ATF6 were not significantly different from those in control group(P>0.05). Compared with JEV group,the apoptosis rate and the expression levels of PERK,CHOP,caspase-12 and Bax in SH-SY5 Y cells of agonist group were significantly increased,and the survival rate was significantly decreased(P<0.05). The apoptosis rate and expression levels of PERK,CHOP,caspase - 12 and Bax in SH -SY5 Y cells of antagonist group were significantly decreased,and the survival rate was significantly increased(P<0.05). Compared with si-NC+JEV group,the apoptosis rate and expression levels of PERK,CHOP,caspase -12 and Bax in SH -SY5 Y cells of si-PERK+JEV group were significantly decreased,the survival rate was significantly increased(P<0.05). These results suggest that the activation of PERK pathway in ERS relates to JEV induced neuronal apoptosis.

关 键 词：流行性乙型脑炎病毒(JEV) 内质网应激 神经元 PERK 凋亡 

分 类 号：R373.31[医药卫生—病原生物学]