miR-140-3p通过靶向趋化因子受体4和抑制JAK2/STAT3通路减轻缺氧复氧诱导的心肌细胞损伤  

miR-140-3p attenuates hypoxia/reoxygenation-induced cardiomyocyte injury by targeting chemokine receptor 4 and inhibiting JAK2/STAT3 pathway

作　　者：蒋芙苓 戴璐 李梦一 JIANG Fuling;DAI Lu;LI Mengyi(Department of Cardiology,Wenzhou Hospital of Traditional Chinese Medicine,Wenzhou,Zhejiang 325000,China;Basic Medical School,Wenzhou Medical University,Wenzhou,Zhejiang 325000,China)

机构地区：[1]温州市中医院心血管内科,浙江省温州市325000 [2]温州医科大学基础医学院,浙江省温州市325000

出　　处：《中国动脉硬化杂志》2022年第9期764-772,共9页Chinese Journal of Arteriosclerosis

摘　　要：[目的]探讨miR-140-3p对缺氧复氧(H/R)诱导的心肌细胞损伤的影响及其机制。[方法]构建体外心肌细胞H/R模型,使用miR-140-3p mimics、趋化因子受体4(CXCR4)过表达质粒转染H9c2细胞。噻唑蓝法检测细胞活性;流式细胞术检测细胞凋亡;反转录聚合酶链反应和Western blot检测细胞中miR-140-3p、CXCR4、Janus蛋白酪氨酸激酶2(JAK2)/信号转导和转录激活因子3(STAT3)通路的激活以及凋亡相关蛋白的表达。双荧光素酶报告基因实验验证miR-140-3p与CXCR4的靶向关系。相应试剂盒检测乳酸脱氢酶(LDH)的活性和炎症因子、活性氧(ROS)的水平。[结果]体外H/R可抑制miR-140-3p的表达,上调CXCR4表达和JAK2/STAT3通路的磷酸化,诱导H9c2细胞凋亡而抑制H9c2细胞增殖,促进炎症因子和ROS的释放并上调LDH的活性。miR-140-3p可以通过靶向CXCR43′UTR抑制CXCR4表达,从而抑制JAK2/STAT3通路的磷酸化激活,抑制炎症因子和ROS的释放,下调LDH的活性,促进H9c2细胞增殖而抑制其凋亡。[结论]miR-140-3p可能通过靶向抑制CXCR4而抑制JAK2/STAT3通路,从而减轻缺血再灌注诱导的心肌细胞损伤。Aim To investigate the effect of miR-140-3p on hypoxia/reoxygenation(H/R)-induced cardiomyocyte injury and its mechanism.Methods In vitro cardiomyocyte H/R model was constructed,and H9c2 cells were transfected with miR-140-3p mimics and chemokine receptor 4(CXCR4)overexpression plasmids.Cell viability was detected by methyl thiazolyl tetrazolium method.Cell apoptosis was detected by flow cytometry.Reverse transcription polymerase chain reaction and Western blot were used to detect miR-140-3p,CXCR4,and the activation of Janus protein tyrosine kinase 2(JAK2)/signal transducers and activators of transcription 3(STAT3)pathway and the expressions of apoptosis-related proteins in cells.The targeting relationship between miR-140-3p and CXCR4 was verified by dual-luciferase reporter gene experiment.The activity of lactate dehydrogenase(LDH)and the levels of inflammatory factors and reactive oxygen species(ROS)were detected with corresponding kits.Results In vitro H/R could inhibit the expression of miR-140-3p,up-regulate the expression of CXCR4 and the phosphorylation of JAK2/STAT3 pathway,induce the apoptosis of H9c2 cells and inhibit the proliferation of H9c2 cells,promote the release of inflammatory factors and ROS,and up-regulate the activity of LDH.miR-140-3p could inhibit the expression of CXCR4 by targeting CXCR43′UTR,thereby inhibiting the phosphorylation activation of JAK2/STAT3 pathway,inhibiting the release of inflammatory factors and ROS,down-regulating the activity of LDH,promoting the proliferation of H9c2 cells and inhibiting their apoptosis.Conclusion miR-140-3p may inhibit the JAK2/STAT3 pathway through targeted inhibition of CXCR4,thereby attenuating ischemia/reperfusion-induced cardiomyocyte injury.

关 键 词：miR-140-3p 缺氧复氧 趋化因子受体4 炎症因子 JAK2/STAT3通路 心肌细胞损伤 

分 类 号：R363[医药卫生—病理学] R5[医药卫生—基础医学]