甲基莲心碱对人脐静脉血管平滑肌细胞增殖及表型调节的影响

Effect of Neferine on the Proliferation and Phenotype of Human Umbilical Vein Smooth Muscle Cells

关键词:  甲基莲心碱  平滑肌细胞  流式细胞术  增殖  表型

Key words:  neferine  smooth muscle cell  flow cytometry  proliferation  phenotype

摘要:

目的 研究甲基莲心碱对人脐静脉平滑肌细胞增殖及表型调节的影响。方法 以0.1，0.5，1.0，5.0 μmol·L^-1甲基莲心碱处理平滑肌细胞，采用MTT 法和流式细胞术观察甲基莲心碱对人脐静脉血管平滑肌细胞增殖的影响。用免疫印迹法检测收缩型平滑肌细胞特异性标志物SM1，calponin 1和α-actin蛋白的表达。结果 甲基莲心碱(0.1~5.0 μmol·L-1)处理12，24，48 h后，人脐静脉血管平滑肌细胞的增殖具有明显的抑制作用，并具有剂量依赖关系和时间依赖关系。在缺乏甲基莲心碱的干预下，血管平滑肌细胞SM1，calponin 1和α-actin蛋白表达减少，在用0.5，5.0 μmol·L^-1甲基莲心碱处理48 h后，可显著逆转SM1，calponin 1和α-actin蛋白表达的减少，且具有剂量依赖性。结论 甲基莲心碱具有抑制血管平滑肌细胞增殖及表型转化的作用，可用于防治动脉粥样硬化和再狭窄。

Abstract:

OBJECTIVE To study the effect of neferine on the proliferation and phenotypic modulation of human umbilical vein smooth muscle cells (HUVSMCs) cultured in vitro. METHODS The effect of neferine(0.1, 0.5, 1.0, 5.0 μmol·L^-1) on HUVSMCs proliferation was studied by using MTT colorimeter and flow cytometry. Western blotting was used to indicate the changes of protein levels of the contractile-phenotype smooth muscle specific markers, such as smooth muscle myosin heavy chain-1 (SM1), calponin 1 and α-actin. RESULTS Neferine could significantly decrease the cell numbers of HUVSMCs after treatment for 12, 24, 48 h, and the anti-proliferation effects were in a concentration-dependent and time-dependent manner when the concentrations of neferine were between 0.1 and 5.0 μmol·L-1. In the absence of neferine treatment, the protein levels of the smooth muscle specific markers (SM1, calponin 1 and α-actin) decreased during culture. However, the treatment of VSMCs with neferine (0.5 or 5.0 μmol·L^-1) for 48 h reversed the effect in a concentration-related manner. CONCLUSION These data suggest that neferine may arrest the growth of VSMCs and inhibit phenotypic modulation, thereby preventing cell dedifferentiation. These properties of neferine suggest that the drug can provide useful therapy for atherosclerosis and restenosis.