丹参酮ⅡA对AngⅡ诱导的大鼠血管平滑肌细胞增殖的影响

Effect of Tanshinone ⅡA on Proliferation of Vascular Smooth Muscle Cell Induced by Angiotensin Ⅱ

目的:探讨丹参酮ⅡA(TSN)对血管紧张素Ⅱ(AngⅡ)诱导的大鼠血管平滑肌细胞增殖的影响,以寻找药物作用的靶点。方法:分离大鼠胸主动脉中层平滑肌,贴壁法培养平滑肌细胞,建立AngⅡ诱导的血管平滑肌细胞(VSMC)增殖模型;以四甲基偶氮唑蓝(MTT)法观察丹参酮ⅡA对VSMC增殖的影响;用酶促反应定磷法测定钙调神经磷酸酶(CaN)活性;应用免疫细胞化学法观察原癌基因c-fos和c-myc的表达。结果:成功建立AngⅡ诱导的VSMC增殖模型;随着TSN浓度增加VSMC增殖活性呈明显下降趋势(P<0.05,0.01);TSN各组随着浓度的增加,VSMC中CaN活性显著下降(P<0.01),VSMC中c-fos和c-myc表达水平也显著下降(P<0.01)。结论:TSN能抑制血管平滑肌细胞的增殖,并呈浓度依赖性。其机制可能与下调VSMC中CaN活性、c-fos及c-myc表达有关。

Objective：To study the effects of tanshinone Ⅱ A （TSN） on proliferation of vascular smooth muscle cells （VSMCs） induced by angiotensin Ⅱ （AngⅡ）. Methods：A cells proliferating model of VSMCs induced by AngⅡ was established. MTT colorimetric assay was used to measure the proliferation of VSMCs. The effects of TSN on CaN were detected by enzyme reaction phosphorus measurement,and the effects of TSN on the c-los and c-myc gene expression exam- ined by immunocytochemical staining and image analysis technique （A value）. Results：The rat aortic SMCs were cultured in vitro successfully. CaN activities,cell proliferation activity and the expression levels of c-fos and c-myc were increased significantly in VSMCs induced by Ang Ⅱ （P〈0.05, P〈0. 01）. However, the indexes above were obviously reduced in TSN treated group as compared with Ang Ⅱ treated group. Different concentrations of TSN （0. 001,0. 01,0.1 mg/L） decreased the CaN activity,cell proliferation activity,and the expression levels of c-fos and c-myc in a dose-dependent manner （P〈0.05 or 0.01）. Conclusion： Ang Ⅱ stimulated the proliferation of VSMCs significantly, which could be inhibited by TSN in a dose-dependent manner by reducing the CaN activity and the expression levels of c-los and c-myc.