MiR-181a-5p通过靶向KLF6信号调控低氧诱导的肺动脉平滑肌细胞的增殖和迁移的机制研究

Mechanism of MiR-181a-5p Regulating Hypoxic-induced Proliferation and Migration of Pulmonary Artery Smooth Muscle Cells by Targeting KLF6 Signaling

目的探讨miR-181a-5p对肺动脉血管平滑肌细胞增殖以及迁移的影响,并探索了相应的分子调控通路。方法本研究于2021年11月—2022年5月进行,实时定量聚合酶链式反应(polymerase chain reaction,PCR)检测肺动脉平滑肌细胞中miR-181a-5p和KLF6 mRNA的表达水平;CCK-8试验检测肺动脉平滑肌细胞的活性;双荧光素酶试验检测miR-181a-5p与KLF6的相互作用。结果低氧诱导能够促进肺动脉平滑肌细胞的活性并抑制miR-181a-5p的表达。miR-181a-5p过表达能够部分反转低氧诱导对肺动脉平滑肌细胞的活性和迁移。miR-181a-5p敲减能够促进肺动脉平滑肌细胞的活性和迁移。miR-181a-5p能够靶向KLF6的3’UTR区域从而抑制KLF6在肺动脉平滑肌细胞的表达。营救试验发现,KLF6过表达能够反转miR-181a-5p过表达对低氧诱导的肺动脉血管平滑肌细胞增殖、迁移的抑制作用。结论本研究结果提示miR-181a-5p能够抑制低氧诱导下的肺动脉平滑肌细胞增殖和迁移,其作用机制可能是通过靶向抑制KLF6表达来实现。

Objective To explore the effects of miR-181a-5p on the proliferation and migration of pulmonary artery vascular smooth muscle cells,and explore the corresponding molecular regulatory pathways.Methods This study was conducted from November 2021 to May 2022.The expression level of miR-181a-5p and KLF6 mRNA in pulmonary artery smooth muscle cells was detected by real-time quantitative polymerase chain reaction(PCR).CCK-8 assay was used to detect the activity of pulmonary artery smooth muscle cells.The interaction between miR-181a-5p and KLF6 was detected by double luciferase assay.Results Hypoxia induction can promote the activity of pulmonary artery smooth muscle cells and inhibit the expression of miR-181a-5p.Overexpression of miR-181a-5p can partially reverse the activity and migration of pulmonary artery smooth muscle cells induced by hypoxia.miR-181a-5p knockdown can promote the activity and migration of pulmonary artery smooth muscle cells.miR-181a-5p can target the 3'UTR region of KLF6,thus inhibiting the expression of KLF6 in pulmonary artery smooth muscle cells.The rescue test found that KLF6 overexpression could reverse the inhibitory effect of miR-181a-5p overexpression on the proliferation and migration of pulmonary artery smooth muscle cells induced by hypoxia.Conclusion The results of this study suggest that miR-181a-5p can inhibit the proliferation and migration of pulmonary artery smooth muscle cells induced by hypoxia,and its mechanism may be achieved through targeted inhibition of KLF6 expression.