Effect of HIF-1α/miR-10b-5p/PTEN on Hypoxia-Induced Cardiomyocyte Apoptosis
Effect of HIF-1α/miR-10b-5p/PTEN on Hypoxia-Induced Cardiomyocyte Apoptosis
复制标题
HIF-1α/miR-10b-5p/PTEN对缺氧诱导的心肌细胞凋亡的影响
DOI：
10.1161/jaha.119.011948
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发表时间：
2019-09-17
影响因子：
5.4
通讯作者：
Cao, Jiumei
中科院分区：
文献类型：
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作者：
Wu, Liping;Chen, Yafen;Cao, Jiumei
背景-很少有报道讨论microRNA miR-10b-5p在低氧条件下调节心肌梗死后(Post-MI)心肌细胞凋亡的机制。方法和结果-C57BL/6小鼠手术结扎左前降支建立MI或缺血/再灌注动物模型。检测不同时间点梗死区组织中miR-10b-5p、PTEN(磷酸酶和张力蛋白同源物)、HIF-1α(低氧诱导因子1α)的表达。胸腔内注射阴性对照或miR-10b-5p后，于1周时在心肌梗死灶周围区域制备高表达慢病毒，检测心肌梗死面积、心功能和心肌细胞凋亡。将miR-10b-5p模拟物导入原代培养的小鼠心肌细胞，分析其对心肌细胞凋亡和PTEN表达的影响。同时，通过荧光素酶报告基因检测证实PTEN是miR-10b-5p的靶标。MiR-10b-5与PTEN共转染证实了miR-10b-5与PTEN的关系。低氧应激条件下，检测HIF-1α和miR-10b-5p的表达。结果显示，miR-10b-5p在梗死区的表达明显降低。在心肌梗死小鼠模型中过表达miR-10b-5p可显著缩小心肌梗死面积，改善心功能，抑制细胞凋亡。在体外过表达miR-10b-5p可拮抗缺氧诱导的心肌细胞凋亡，并特异性地抑制凋亡相关基因PTEN的表达，但过表达PTEN可减弱这些作用。我们还发现，缺氧诱导的HIF-1α的积聚导致miR-10b-5p的表达减少。干扰HIF-1α信号通路的激活促进了PrI-miR-10b和miR-10b-5p的表达，抑制了PTEN的表达。结论microRNA miR-10b-5p可拮抗缺氧诱导的心肌细胞凋亡，提示miR-10b-5p可能成为治疗MI的潜在临床靶点。
Background-Few reports have addressed the mechanism by which microRNA miR-10b-5p regulates post-myocardial infarction (post-MI) cardiomyocyte apoptosis under hypoxic conditions.Methods and Results-C57BL/6 mice underwent surgical ligation of the left anterior descending artery to create an MI or ischemia/reperfusion animal model. The expression of miR-10b-5p, PTEN (phosphatase and tensin homolog), and HIF-1 alpha (hypoxiainducible factor 1 alpha) was detected in infarct border zone tissues at various time points. After precordial injections of the negative control or miR-10b-5p, overexpression lentiviruses were made in the areas surrounding the MI sites at 1 week, and myocardial infarct size, cardiac function, and cardiomyocyte apoptosis were examined. A miR-10b-5p mimic was transfected into primary mouse cardiomyocytes to analyze its effects on cardiomyocyte apoptosis and PTEN expression. Meanwhile, PTEN as a target of miR-10b-5p was verified via luciferase reporter gene assays. Cotransfection of miR-10b-5 and PTEN verified the relationship between miR-10b-5 and PTEN. Under hypoxic stress, the expression of HIF-1 alpha and miR-10b-5p was examined. The results showed that miR-10b-5p expression was markedly reduced in the infarct border zone. Overexpression of miR-10b-5p in the murine model of MI significantly reduced MI size, improved cardiac function, and inhibited apoptosis. Overexpression of miR-10b-5p in vitro antagonized hypoxia-induced cardiomyocyte apoptosis and specifically inhibited the expression of the apoptosis-related gene PTEN, but overexpression of PTEN weakened these effects. We also found that hypoxia-induced accumulation of HIF-1 alpha resulted in decreased expression of miR-10b-5p. Interfering with the activation of the HIF-1 alpha signaling pathway promoted Pri-miR-10b and miR-10b-5p expression and inhibited PTEN expression.Conclusions-MicroRNA miR-10b-5p antagonizes hypoxia-induced cardiomyocyte apoptosis, indicating that miR-10b-5p may serve as a potential future clinical target for the treatment of MI.