Department of Cardiovascular Medicine, The First Affiliated Hospital of Bengbu Medical College, Bengbu 233004, Anhui, China.
J Biosci. 2020;45.
This paper explores the potential mechanism of microRNA-143-5p regulation effects on pulmonary artery smooth muscle cells (PASMCs) functions in hypoxic pulmonary hypertension (HPH) via targeting HIF-1a, which may offer a new idea for HPH therapy. PASMCs were transfected with mimics control/miR-143-5p mimics or inhibitor control/miR-143-5p inhibitor. We used Western blotting and RT-qPCR to detect the protein and mRNA expressions, CCK-8 assay to detect cellular viability, Annexin V-FITC/PI staining and caspase- 3/cleaved caspase-3 protein to evaluate cellular apoptosis, transwell migration experiment for cellular migration measurement and Dual luciferase reporter gene assay to prove the target of miR-143-5p. Cells under hypoxic condition presented the decreased protein and mRNA expressions of α-smooth muscle actin (SM-α-actin), Myocardin, smooth muscle myosin heavy chain (SMMHC), and smooth muscle-22α (SM22α), Calponin1 and Hypoxia-inducible factor-1α(HIF-1α), the increased cell viability and miR-143-5p level; Overexpression of miR-143-5p obviously reduced vascular smooth muscle-specific contraction marker protein levels and cellular apoptosis, increased cellular migration of PASMCs with hypoxia stimulation; Low-expression of miR-143-5p caused the opposite changes, while co-transfected with Si HIF-1 α blocked the beneficial effects of miR-143-5p inhibition on PASMCs under hypoxia. MicroRNA-143-5p can promote the phenotype conversion, proliferation and migration of pulmonary artery smooth muscle cells under hypoxic condition through direct targeting of HIF-1α.
本文探讨了 microRNA-143-5p 通过靶向 HIF-1a 对肺动脉平滑肌细胞(PASMC)功能的潜在调节作用机制在低氧性肺动脉高压(HPH)中的作用,这可能为 HPH 治疗提供新的思路。转染 mimics 对照/miR-143-5p mimics 或 inhibitor 对照/miR-143-5p inhibitor 到 PASMCs 中。我们使用 Western blot 和 RT-qPCR 检测蛋白和 mRNA 表达,CCK-8 检测细胞活力,Annexin V-FITC/PI 染色和 caspase-3/cleaved caspase-3 蛋白检测细胞凋亡,Transwell 迁移实验检测细胞迁移,双荧光素酶报告基因实验验证 miR-143-5p 的靶基因。低氧条件下的细胞表现出 α-平滑肌肌动蛋白(SM-α-actin)、肌球蛋白重链(Myocardin)、平滑肌肌球蛋白重链(SMMHC)和平滑肌-22α(SM22α)、钙调蛋白 1 和低氧诱导因子-1α(HIF-1α)的蛋白和 mRNA 表达降低,细胞活力和 miR-143-5p 水平升高;miR-143-5p 的过表达明显降低了血管平滑肌特异性收缩标志物蛋白水平和细胞凋亡,增加了低氧刺激下 PASMCs 的细胞迁移;miR-143-5p 的低表达则引起相反的变化,而共转染 Si HIF-1α 则阻断了 miR-143-5p 抑制对低氧下 PASMCs 的有益作用。miR-143-5p 可通过直接靶向 HIF-1α,促进低氧条件下肺动脉平滑肌细胞的表型转化、增殖和迁移。