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赛培新增产品文章：大鼠TNF-α、IL-1β、IL- 6、IL- 10、TGF-β
发布时间：2022-07-25 11:25:45
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
蒋芙苓1，戴 璐2，李梦一2 ( 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) ［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 ［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 CXCR4 3'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。 ［中图分类号］ R363; R5 ［文献标识码］ A miR-140-3p attenuates hypoxia /reoxygenation-induced cardiomyocyte injury by tar- geting chemokine receptor 4 and inhibiting JAK2 /STAT3 pathway JIANG Fuling1，DAI Lu2，LI Mengyi2 ( 1． Department of Cardiology，Wenzhou Hospital of Traditional Chinese Medicine，Wenzhou，Zhejiang 325000，China; 2． Basic Medical School，Wenzhou Medical University，Wenzhou，Zhejiang 325000，China) ［KEY WORDS］ miR-140-3p; hypoxia /reoxygenation; chemokine receptor 4; inflammatory factor; JAK2 / STAT3 pathway; cardiomyocyte injury ［ABSTRACT］ Aim To investigate the effect of miR-140-3p on hypoxia /reoxygenation ( H/R) -induced cardiomyo- cyte 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 de- tected by methyl thiazolyl tetrazolium method． Cell apoptosis was detected by flow cytometry． Reverse transcription pol- ymerase chain reaction and Western blot were used to detect miR-140-3p，CXCR4，and the activation of Janus protein tyro- sine kinase 2 ( JAK2) /signal transducers and activators of transcription 3 ( STAT3) pathway and the expressions of apopto- sis-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-reg- ulate the activity of LDH． miR-140-3p could inhibit the expression of CXCR4 by targeting CXCR4 3'UTR，thereby inhib- iting 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通过靶...缺氧复氧诱导的心肌细胞损伤_蒋芙苓.pdf下载
