丹参酮ⅡA对血管紧张肽Ⅱ诱导增殖的大鼠血管平滑肌细胞中c-fos与c-myc的影响

Tanshinone ⅡA Represses c-fos、c-myc Expression of the Proliferating Vascular Smooth Muscle Cells Activated by Angiotensin Ⅱ

目的探讨丹参酮ⅡA对血管紧张肽Ⅱ(AngⅡ)诱导的大鼠血管平滑肌细胞(VSMC)增殖的影响及其机制。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;流式细胞仪检测药物对细胞周期的影响;应用免疫细胞化学方法观察原癌基因c-fos和c-myc表达水平的变化。结果成功建立AngⅡ诱导的VSMC增殖模型;随着丹参酮ⅡA浓度增加,VSMC增殖活性呈明显下降趋势(P<0.05);流式细胞术结果显示丹参酮ⅡA使VSMC G0/G1期比例升高(P<0.01),S期比例下降(P<0.01);丹参酮ⅡA各组随着浓度的增加,VSMC中c-fos和c-myc表达水平显著下降(P<0.05)。结论丹参酮ⅡA具有抑制血管平滑肌细胞增殖的作用,并呈浓度依赖性。其机制可能与丹参酮ⅡA下调VSMC中c-fos及c-myc表达水平有关。

Objective To study the effects of tanshinone Ⅱ A （TSN） to c-los and c-myc of the proliferating vascular smooth muscle cell（VSMC） treated by angiotensin Ⅱ （Ang Ⅱ ）. Methods A cell proliferating model of VSMC induced by angiotensin Ⅱ was established and then measured by MTT colorimetric assay. The effects of TSN on cell cycle of VSMC was detected by flow cytometry and on c-los and c-myc gene expression by immunocytochemical staining （A value）. Results The rat aortic smooth muscle cells were cultured in vitro successfully. The proportion of VSMC G0/G1 period was ascended by TSN （P 〈 0.01 ） and the proportion of VSMC S period was descended（ P 〈 0.01 ）. Cell proliferation activity and the expression levels of c-los and c-myc were increased significantly in proliferating VSMC induced by Ang Ⅱ （ P 〈 0.01 ）. While treated with TSN, the indexes were obviously reduced in a dose-dependent manner （0. 001,0. 010 ,0. 100 mg · L^-1 ） （ P 〈 0.05 ,P 〈 0.01 ）. Conclusion Ang Ⅱ significantly stimulated the proliferation of VSMC. The VSMC proliferation induced by Ang Ⅱ can be inhibited by TSN in dose-dependent manner and the mechanism may be that TSN decreases the expression levels of c-los and c- myc.