氟西汀通过RhoA信号通路抑制5-羟色胺诱导的肺动脉平滑肌细胞增殖

Fluxetine Inhibited Serotonin-induced Pulmonary Arterial Smooth Muscle Cells Proliferation via Rho A Signalling Pathway

目的研究氟西汀对5-羟色胺(5-HT)诱导的肺动脉平滑肌细胞(PASMC)Rho A信号通路的影响。方法培养大鼠PASMC,用氟西汀(0.1、1、10μmol/L)或法舒地尔(1、10、100μmol/L)干预后,加入1μmol/L 5-HT刺激48 h,以噻唑蓝比色法观察细胞增殖情况,以免疫共沉淀、Western blot等检测Rho A信号通路各指标。结果5-HT刺激PASMC增殖,使Rho A 5-HT化、Rho A膜转位、Rho激酶蛋白2表达以及肌球蛋白磷酸酶目标亚基1、细胞外调节蛋白激酶、蛋白激酶B磷酸化明显增加;氟西汀剂量依赖地抑制这些改变;而Rho激酶抑制剂法舒地尔对Rho A 5-HT化没有抑制作用。结论氟西汀通过阻断Rho A信号通路抑制5-HT诱导的PASMC增殖。

Aim To study the effect of fluoxetine on serotonin-induced Rho A signalling pathway in pulmonary arterial smooth muscle cells. Methods The pulmonary arterial smooth muscle cells of rat were exposed to serotonin for48 h with or without fluoxetine or fasudil. Then pulmonary arterial smooth muscle cells proliferation was tested by 3-[4,5-dimethylthylthiazol-2-yl]-2,5 diphenyltetrazolium broide assay. The cellular extracts were prepared for coimmunoprecipitation or Western blot of Rho A signalling pathway. Results Serotonin promoted pulmonary arterial smooth muscle cells proliferation,increased Rho A serotonylation,Rho A membrane translocation,Rho kinase 2 protein expression and myosin phosphatase target subunit 1,extracellular regulated protein kinase and protein kinase B phosphorylation. Fluoxetine inhibited these changes dose-dependently. However,fasudil did not inhibit increased Rho A serotonylation induced by serotonin. Conclusion Fluxetine inhibited serotonin-induced pulmonary arterial smooth muscle cells proliferation via inhibition of Rho A signalling pathway.