Nrf2/HO-1通路通过调控内质网应激改善小鼠心肌细胞缺氧/复氧损伤的作用研究  

Nrf2/HO-1 pathway improves anoxia/reoxygenation-induced injury in murine cardiomyocytes via regulating endoplasmic reticulum stress

作　　者：陶卉 段晓宇[2] 刘筱 张燕[1] 田晓芬 吴群英 于杨 TAO Hui;DUAN Xiao-yu;LIU Xiao;ZHANG Yan;TIAN Xiao-fen;WU Qun-ying;YU Yang(Department of Cardiac Function,The Central Hospital of Wuhan,Tongji Medical College,Huazhong University of Science and Technology,Wuhan Hubei 430014,China;Department of General Medicine,The Central Hospital of Wuhan,Tongji Medical College,Huazhong University of Science and Technology,Wuhan Hubei 430014,China)

机构地区：[1]华中科技大学同济医学院附属武汉市中心医院心功能科,湖北武汉430014 [2]华中科技大学同济医学院附属武汉市中心医院全科医学科,湖北武汉430014

出　　处：《蚌埠医学院学报》2023年第3期296-300,共5页Journal of Bengbu Medical College

基　　金：湖北省武汉市卫健委科研课题(WZ17D02)。

摘　　要：目的:探讨Nrf2/HO-1信号上调对小鼠心肌细胞缺氧/复氧损伤的改善作用及相关机制。方法:小鼠原代心肌细胞分为正常培养处理和缺氧/复氧处理,其中缺氧/复氧处理的心肌细胞分为模型组、Nrf2过表达组、HO-1过表达组、HO-1沉默+Nrf2过表达组。采用MTT法检测细胞活性,TUNEL染色检测细胞凋亡,ELISA法检测细胞培养上清中心肌肌钙蛋白Ⅰ(cTnⅠ)及肌酸激酶同工酶(CK-MB)含量,荧光定量PCR检测内质网应激相关分子水平。结果:小鼠心肌细胞缺氧/复氧处理后,心肌细胞活性降低、细胞凋亡增加及细胞培养上清中cTnⅠ和CK-MB含量升高(P<0.05)。Nrf2或HO-1过表达可有效减轻缺氧/复氧诱导的心肌细胞损伤(P<0.05)。给予HO-1基因沉默预处理后,Nrf2过表达产生的细胞保护作用明显减弱(P<0.05)。此外,缺氧/复氧处理后,心肌细胞内GRP78、ATF6、CHOP和Caspase-3的mRNA表达量升高(P<0.05)。Nrf2或HO-1过表达引起GRP78、ATF6、CHOP和Caspase-3表达水平下调(P<0.05),而下调HO-1水平可使Nrf2过表达对内质网应激信号转导的抑制作用减弱(P<0.05)。结论:Nrf2/HO-1通路表达上调可有效改善缺氧/复氧诱导的心肌细胞损伤,其机制可能在于抑制内质网应激以及下游的细胞凋亡通路信号转导。Objective:To discuss the effect of Nrf2/HO-1 up-regulation on improving anoxia/reoxygenation-induced injury in murine cardiomyocytes and its related mechanism.Methods:The primary murine cardiomyocytes were divided into normal culture treatment and anoxia/reoxygenation treatment,and the anoxia/reoxygenation treated cardiomyocytes were divided into model group,Nrf2 overexpression group,HO-1 overexpression group,HO-1 silence+Nrf2 overexpression group.Cell viability was detected by MTT assay,apoptosis was analyzed by TUNEL staining,cardiac troponinⅠ(cTnⅠ)and creatine kinase isoenzyme MB(CK-MB)contents in cell culture supernatant were determined by ELISA,and levels of endoplasmic reticulum stress-related molecules were measured by fluorescence quantitative PCR.Results:After anoxia/reoxygenation treatment,the activity decreased,apoptosis increased,and the contents of cTnⅠand CK-MB in cell culture supernatant of murine cardiomyocytes increased(P<0.05).Overexpression of Nrf2 or HO-1 could effectively alleviate anoxia/reoxygenation-induced injury in murine cardiomyocytes(P<0.05).After pretreatment with HO-1 gene silencing,the protection effect of Nrf2 overexpression was significantly weakened(P<0.05).In addition,after anoxia/reoxygenation treatment,the mRNA expressions of GRP78,ATF6,CHOP and Caspase-3 in cardiomyocytes increased(P<0.05).Overexpression of Nrf2 or HO-1 resulted in down-regulation of the expression levels of GRP78,ATF6,CHOP and Caspase-3(P<0.05),while down-regulation of HO-1 level reduced the inhibitory effect of Nrf2 overexpression on endoplasmic reticulum stress signal transduction(P<0.05).Conclusions:The up-regulation of Nrf2/HO-1 pathway can effectively improve anoxia/reoxygenation-induced injury in cardiomyocytes,which may be related to the suppression of endoplasmic reticulum stress and its downstream apoptosis signaling transduction.

关 键 词：急性心肌梗死 缺氧/复氧损伤 内质网应激 Nrf2/HO-1通路 

分 类 号：R542.2[医药卫生—心血管疾病]