Jia Qihua, Li Ling, Song Wenjie, Cao Nan, Li Li, Ma Ketao, Si Junqiang
Ministry-of-Education Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Physiology, School of Medicine, Shihezi University, Shihezi 832000, China.
Department of Traumatology, First Affiliated Hospital, Shihezi University, Shihezi 832002, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2020 Jul;36(7):616-621.
Objective To explore the role of Cx43 in the proliferation and migration of human pulmonary arterial smooth muscle cells (HPASMCs) induced by angiotensin II (Ang II). Methods HPASMCs were cultured in vitro and randomly divided into four groups: control group, Ang II group, Ang II combined with DMSO group, and Ang II combined with candesartan group, and cells were collected in logarithmic growth phase. Cell viability was detected by CCK-8 assay; the migration ability of HPASMCs were measured by wound-healing and Transwell assay. The protein levels of Cx43, osteopontin (OPN), proliferating cell nuclear antigen (PCNA), SMAD2 and SMAD3 in HPASMCs were detected by Western blot analysis. Results Compared with the control group, the expression of OPN and PCNA proteins significantly went up in Ang II group, and the cell proliferation and migration ability increased. The cell proliferation and migration ability of the Ang II combined with candesartan group were significantly lower than that in the Ang II group. Compared with the control group, the Cx43 protein and its phosphorylation level increased significantly in the Ang II group, and the protein expression of SMAD2 and SMAD3 increased, while the expression of each protein in the Ang II combined with candesartan group was significantly lower than those in the Ang II group. Conclusion Ang II up-regulates the expression of Cx43 protein to promote the proliferation and migration of HPASMCs, which may be related to the activation of SMAD2/3 signaling pathway.
目的 探讨Cx43在血管紧张素II（Ang II）诱导的人肺动脉平滑肌细胞（HPASMCs）增殖和迁移中的作用。方法 体外培养HPASMCs，随机分为四组：对照组、Ang II组、Ang II联合二甲基亚砜（DMSO）组、Ang II联合坎地沙坦组，收集对数生长期细胞。采用CCK-8法检测细胞活力；采用划痕实验和Transwell实验检测HPASMCs的迁移能力。通过蛋白质印迹法检测HPASMCs中Cx43、骨桥蛋白（OPN）、增殖细胞核抗原（PCNA）、SMAD2和SMAD3的蛋白水平。结果 与对照组相比，Ang II组OPN和PCNA蛋白表达显著升高，细胞增殖和迁移能力增强。Ang II联合坎地沙坦组的细胞增殖和迁移能力显著低于Ang II组。与对照组相比，Ang II组Cx43蛋白及其磷酸化水平显著升高，SMAD2和SMAD3蛋白表达增加，而Ang II联合坎地沙坦组各蛋白表达均显著低于Ang II组。结论 Ang II上调Cx43蛋白表达以促进HPASMCs的增殖和迁移，这可能与SMAD2/3信号通路的激活有关。
