miR-21-5p在肺动脉高压发病机制中的调控作用研究

Study of regulating effect of mi R-21-5p in pulmonary arterial hypertention pathogenesis

目的研究mi R-21-5p在肺动脉高压发病中的调控机制。方法应用q RT-PCR方法检测mi R-21-5p在正常人和先天性心脏病伴肺动脉高压患者血清,以及常氧和缺氧处理肺动脉平滑肌细胞中的表达;利用Edu渗入法以及划痕实验研究mi R-21-5p对肺动脉平滑肌细胞增殖及迁移的影响;应用Targetscan软件进行靶基因预测并在肺动脉平滑肌细胞中进行验证。结果肺动脉高压患者血清mi R-21-5p水平显著高于正常人血清(P<0.001);与常氧相比,缺氧处理肺动脉平滑肌细胞明显促进mi R-21-5p表达(P<0.01);mi R-21-5p能够促进缺氧条件下的肺动脉平滑肌细胞增殖、迁移;双荧光素酶报告系统证明mi R-21-5p能够结合骨形成蛋白Ⅱ型受体(BMPR2)的3'UTR;鉴定mi R-21-5p在肺动脉平滑肌细胞中靶向BMPR2基因。结论 mi R-21-5p调控靶基因BMPR2参与肺动脉高压发病。

Objective To reveal the role of mi R-21-5p in the development of pulmonary arterial hypertention(PAH).Methods q RT-PCR method was used to detect the expression of mi R-21-5p in the serum of healthy donors and patients with PAH associated with congenital heart disease(CHD-PAH), and the pulmonary artery smooth muscle cells(PASMC) exposed to normoxia and hypoxia. The effects of mi R-21-5p on hypoxia-induced PASMC proliferation and migration were detected by Edu incorporation and wound assay. The target gene of mi R-21-5p was predicted by targetscan and validated by luciferase report assay. Results mi R-21-5p level was significantly increased in the serum of CHD-PAH patient as compared with healthy donors(P < 0.001). In addition, the expression of mi R-21-5p was significantly induced in PASMC treated by hypoxic as compared with normoxia(P < 0.01). Functional analysis revealed that mi R-21-5p significantly enhanced hypoxia-induced PASMC proliferation and migration. Finally, dual-luciferase reporter system proved mi R-21-5p could integrate with BMPR2 3'UTR, then BMPR2 was identified as a direct target of mi R-21-5p. Conclusion The study demonstrates that mi R-21-5p is involved in regulating PAH by targeting BMPR2.