AMP依赖的蛋白激酶α2介导二十二碳六烯酸对抗血管平滑肌细胞表型转化研究

AMPK α2 mediated the beneficial effects of DHA on VSMCs phenotypic switch

目的探讨AMP依赖的蛋白激酶(AMPK)α2介导二十二碳六烯酸(DHA)对抗血管平滑肌细胞表型转化的调控作用。方法原代培养小鼠血管平滑肌细胞(VSMC),应用棕榈酸(PA)做为平滑肌细胞表型转换的体外病理刺激因素,应用不同浓度的DHA进行干预,流式细胞术检测细胞的增殖情况;明胶谱分析、Transwell小室观察细胞的迁移情况;Western blot检测平滑肌肌球蛋白重链(SMMHC)表达及细胞分化情况,同时检测AMPKα表达及活化情况。结果 PA处理后,原代小鼠VSMC增殖能力显著增强,G1向G2细胞周期转化的细胞比例增加;DHA作用后,PA诱导的VSMC增殖能力明显受到抑制。明胶酶谱分析和Transwell小室研究发现,DHA处理后,PA诱导的VSMC的迁移能力受到明显抑制;细胞增殖标志蛋白增殖细胞核抗原(PCNA)表达下降,细胞分化标志蛋白SMMHC表达增加,DHA可剂量依赖性抑制PA诱导的VSMC表型转换。DHA可诱导p AMPKα表达,且以AMPKα2异构体为主。结论 AMPKα2异构体介导了DHA对抗PA诱导的血管平滑肌细胞表型转化。

Objective To investigate the regulatory effect of AMP-dependent protein kinase（AMPK） second mediated docosahexaenoic acid（DHA） on vascular smooth muscle cell phenotypic transformation.Methods Primary cultured mouse vascular smooth muscle cells（VSMC） were used as the in vitro pathological stimulator of smooth muscle cell phenotype conversion using palmitic acid（PA）,different concentrations of DHA were used for intervention,and cell proliferation was detected by flow cytometry.Gelatin spectrum analysis and Transwell cell observation of cell migration.The expression and cell differentiation of smooth muscle myosin heavy chain（SMMHC）and the activation of AMPKα were detected by Western blot. Results After PA treatment,the proliferation capacity of VSMC in primary mice was significantly enhanced,and the proportion of cells transformed from G1 to G2 cell cycle increased.The PA-induced proliferation of VSMC was significantly inhibited by DHA.After DHA treatment,PA-induced migration of VSMC was significantly inhibited.The expression of cell proliferation marker nuclear antigen（PCNA） reduced,the expression of cell differentiation marker SMMHC increased,and DHA could inhibit PA-induced VSMC phenotype conversion in a dose-dependent manner.DHA could induce pAMPKα expression,and AMPKα2 isomer is the main one.Conclusion AMPKα2 mediates the PA-induced phenotypic transformation of DHA in vascular smooth muscle cells.