从CaN信号通路探讨川芎嗪对Ang Ⅱ诱导的大鼠VSMCs增殖的抑制作用及机制  

Effects of Ligustrazine on Inhibiting VSMCs Proliferation Induced by Angiotensin Ⅱ Through Calcineurin Signal Pathway

作　　者：李映红[1] 吴正治 郑红花[1] 罗德生[1] 吴伟康[3] 

机构地区：[1]咸宁学院医学院生物化学与分子生物学系,湖北咸宁437100 [2]广东省深圳市中西医结合临床研究所,广东深圳518035 [3]中山大学基础医学院,广东广州510080

出　　处：《上海中医药杂志》2006年第5期46-49,共4页Shanghai Journal of Traditional Chinese Medicine

基　　金：湖北省教育厅重点科研基金资助项目(2003A006)

摘　　要：目的从钙调神经磷酸酶(CaN)信号通路探讨川芎嗪对血管紧张素Ⅱ(AngⅡ)诱导的大鼠血管平滑肌细胞(VSMCs)增殖的抑制作用及机制。方法以AngⅡ诱导体外培养的大鼠主动脉平滑肌细胞建立细胞增殖模型,实验分为空白组、AngⅡ组、AngⅡ+川芎嗪(低、中、高)剂量组。采用定磷法分别测定各组细胞CaM和CaN活性,MTT法检测细胞增殖活度的变化,免疫组化定量技术观察细胞内c-myc、PCNA的表达水平。结果AngⅡ组VSMCs细胞增殖活度(吸光度值)、胞内CaM、CaN活性以及c-myc、PCNA表达量(光密度值)显著高于空白组(P<0.01);同时加入不同剂量(低、中、高剂量)川芎嗪温育,各组CaM、CaN活性以及c-myc、PCNA表达水平均显著低于AngⅡ组(P<0.05或P<0.01)。结论川芎嗪可通过降低细胞内CaM、CaN活性,下调胞内c-myc、PCNA表达水平,从而显著抑制AngⅡ诱导的VSMCs增殖。Objective To explore the inhibiting effects of Ligustrazine on the proliferation of vascular smooth muscle cells （VSMCs） induced by angiotensin Ⅱ（AngⅡ）and study its mechanism from calcineurin signal pathway. Methods Cultured cells proliferating model of rats＇VSMCs were established by AngⅡ. There were five groups：control group,AngⅡ group,Ang Ⅱ and Ligustrazine （low dose,middle dose,high dose）respectively. Calmodulin（CaM）and calcineurin（CaN）activities were determined by enzyme reaction phosphorus measurement. MTT colorimetric assay was used to measure the proliferation of VSMCs. The expression of intranuclear proto-oncogenes c-myc and proliferating cell nuclear antigen （PCNA）were observed by immunocytoehemical stain and imageanalysis technique. Results AngⅡ increased the proliferation activity, CaM or CaN activities and the expression of c-myc or PCNA with marked statistical difference comparing with the control group（P〈 0.05 or P〈 0.01）. While the VSMCs were incubated with Ligustrazine in different doses[low dose（40 mg /L）,middle dose （400 mg /L）, high dose （4000 mg /L）] at the same time, the indexes above were obviously reduced comparing with AngⅡ group （P〈 0.05 or P〈 0.01）. Conclusion The proliferation of rat VSMCs induced by AngⅡ can be inhibited by Ligustrazine significantly,and the inhibiting mechanism of Ligustrazine may be related to inhibiting CaM or CaN activities, and then restraining the expressions of c-myc or PCNA in a dose and time-dependent manner.

关 键 词：川芎嗪 药理学 血管平滑肌细胞 钙调神经磷酸酶 增殖 

分 类 号：R285[医药卫生—中药学]