碘化N-正丁基氟哌啶醇抑制机械损伤诱导的血管平滑肌细胞增殖的作用及机制

Effect and mechanism of N-n-butyl haloperidol iodide on the proliferation of rat vascular smooth muscle cells induced by mechanical injury

目的探讨碘化N-正丁基氟哌啶醇(F2)对机械损伤所致血管平滑肌细胞(VSMCs)增殖的抑制作用及其机制。方法原代培养大鼠胸主动脉VSMCs,体外建立机械划伤致VSMCs增殖模型,采用Cell Counting Kit-8(CCK-8)法检测细胞增殖情况;流式细胞术法测定细胞周期;Western blot法检测p-MEK,p-ERK,Egr-1蛋白表达;Real Time RT-PCR检测MEK和Egr-1 mRNA表达。结果 F2剂量依赖地抑制机械损伤所致VSMCs增殖,降低损伤刺激下VSMCs的生存率和DNA合成,使G0/G1期细胞比例升高,G2/M期细胞百分比下降,并且抑制损伤后VSMCs中MEK/ERK的活化,使p-MEK/ERK蛋白表达降低,同时下调损伤后MEK和Egr-1的高表达。结论 F2对机械损伤所致血管平滑肌细胞增殖有明显抑制作用,其机制可能与影响MEK/ERK/Egr-1信号途径有关。

Aim To investigate the inhibitory effects and mechanisms of N-n-butyl haloperidol iodide （F2 ） on rat vascular smooth muscle cells（VSMCs） prolifera- tion induced by mechanical injury in vitro. Methods The primarily cultured rat thoracic aortic SMCs were used to establish mechanical injury models in vitro. The proliferation activity of VSMCs was analyzed by cell counting Kit-8 （CCK-8） assay. And the cell cycle was analyzed by flow cytometry. The levels of Egr-1 and p-ERK, p-MEK were assessed by Western blot. The expressions of MEK and Egr-1 mRNA were tested by real time RT-PCR. Results F2 concentration-de- pendently inhibited VSMCs proliferation, decreased the cell survival rate and DNA synthesis in VSMCs stimulated by mechanical injury, which led to the increase of cell numbers of G0/G1 phase and a reduction in those of GJM phase, and restrained the activation of MEK/ERK by attenuating phospho-MEK/ERK and Egr-1 proteins expression as well as down-regulated the overexpression of MEK and Egr-1 after mechanical injury. Conclusion F2 obviously inhibits vascular smooth muscle cell proliferation after mechanical injury in vitro, and its mechanisms may be involved in inhibi- ting the MEK/ERK/Egr-1 signaling pathway.