川芎嗪对Ang Ⅱ诱导的大鼠VSMCs增殖的干预作用

Interference effects of tetramethylpyrazine on the proliferation of vascular smooth muscle cells(VSMCs) induced by angiotensin Ⅱ

目的探讨不同浓度川芎嗪不同作用时间对血管紧张素Ⅱ(AngⅡ)诱导的血管平滑肌细胞(VSMCs)增殖的影响。方法建立AngⅡ诱导VSMCs增殖模型,用酶促反应定磷法观察不同浓度川芎嗪在不同作用时段对血管平滑肌细胞CaN活性的影响;应用免疫细胞化学法观察原癌基因c-fos和增殖细胞核抗原(PCNA)表达水平的变化。结果成功建立了AngⅡ诱导的VSMCs增殖模型,与对照组比较,AngⅡ组刺激后VSMCs增殖活度明显升高(P<0·05);川芎嗪各组CaN活性和c-fos及PCNA表达水平随川芎嗪的浓度和作用时间的延长而显著下降(P<0·05)。结论AngⅡ能显著刺激大鼠血管平滑肌细胞增殖,川芎嗪能使血管平滑肌细胞中PCNA和原癌基因c-fos的表达减弱,其抑制细胞增殖的作用机制可能与其干预CaN依赖的信号转导途径有关,且在一定范围内呈剂量和时间依赖性。

Objective To study the interference effects of tetramethylpyrazine （TMP） on calcinuerin （CAN）, c-fos and the nuclear antigen of proliferating cells in the proliferation of vascular smooth muscle cells （VSMCs） treated by angiotensin Ⅱ （Ang Ⅱ）. Methods A cell proliferating model of VSMCs induced by Ang Ⅱ was established; the effects of TMP on CAN was detected by enzyme reaction phosphorus measurement; the effects of TMP on c-fos gene and PCNA expression were observed by irnmunocytochemical staining and image analysis technique （A value）. Results The rats＇ aortic smooth muscle cells were cultured in vitro successfully. CaN activities, cell proliferation activity and the expression levels of c-fos and PCNA increased significantly in VSMCs proliferation induced by Ang Ⅱ （P〈0.05）. While treated with TMP, the indexes were obviously reduced compared with Ang Ⅱ group. Different concentrations of TMP （40μg/ml, 400μg/ml, 4000μg/ml） decreased the CaN activity and the expression levels of c-fos and PCNA in a dose-dependent manner. Different time of the same dose of TMP also reduced markedly CaN activity and the expression levels of c-los and PCNA in a time-dependent manner. All of them were obviously reduced in a dose and time-dependent manner while treated with TMP （P〈0.05）. Conclusions Ang Ⅱ significantly stimulated the proliferation of VSMCs. The VSMCs proliferation induced by Ang Ⅱ can be inhibited by TMP in a dose and time-dependent manner and the inhibiting mechanism of TMP may be related to inhibiting CaN activities and the expression of c-fos and PCNA.