摘要
目的研究川芎嗪抑制血管平滑肌细胞(vascular smooth muscle cells,VSMCs)增殖的作用机制,为中药有效预防血管再狭窄提供实验依据。方法体外培养大鼠胸主动脉VSMCs,分为:阴性对照组、血管紧张素Ⅱ(AngⅡ)组(AngⅡ10^(-6) mol/L)、低剂量组(川芎嗪20μmol/L加AngⅡ]、中剂量组(川芎嗪40μmol/L加AngⅡ)及高剂量组(川芎嗪80μmol/L加AngⅡ)。MTT法检测细胞增殖率;Western blot和实时荧光定量PCR法分别检测Wnt通路相关蛋白Wnt4、β-catenin、Dvl-1、CyclinD1及胶原Ⅰ(ColⅠ)、胶原Ⅲ(ColⅢ)蛋白、基因表达。结果与阴性对照组比较,AngⅡ组细胞增殖率明显升高(P<0.05)。与AngⅡ组比较,中、高剂量组细胞增殖率明显降低(P<0.05)。与阴性对照组比较,AngⅡ组Wnt4、β-catenin、Dvl-1、CyclinD1、ColⅠ、ColⅢ蛋白和mRNA表达均明显上调(P<0.05)。与AngⅡ组比较,中、高剂量组Wnt4、β-catenin、Dvl-1、CyclinD1、ColⅠ、ColⅢ蛋白和mRNA表达均明显下调(P<0.05)。上述指标在低、中、高剂量组均具有浓度依赖性。结论川芎嗪通过下调Wnt信号通路在一定程度上抑制了AngⅡ诱导的VSMCs的增殖及胶原分泌。
Objective To study the action mechanism of tetramethylpyrazine （TMP） on the pro- liferation of vascular smooth muscle cells （VSMCs）, thus providing experimental evidence for Chinese medicing to effectively prevent restenosis. Methods Rats＇ thoracic aorta VSMCs in vitro cultured （cell line A7r5） were divided into five groups, i.e., the negative control group, the angiotensin II （Ang II , 10-6mol/L） group, the low dose TMP （20 μmol/L） plus Ang IIgroup, the middle dose TMP （40 μmol/L） plus Ang II group, the high dose TMP （80 μmol/L） plus Ang II group. The proliferation ratio was detected by Mtt. Gene and protein expressions of Wnt4, Dvl-1, beta-catenin, CyclinD1, and collagen I and III were detected with real-time fluorescent quantitative PCR and Western blot respectively. Results Com- pared with the negative control group, the proliferation ratio of VSMCs obviously increased in the Ang II group （P 〈0.05）. Compared with the AngII group, the proliferation ratio of VSMCs obviously decreased in the middle dose TMP plus Ang II group and the high dose TMP plus Ang IIgroup （P 〈0.05）. Com- pared with the negative control group, gene and protein expressions of Wnt4, Dvl-1, 13-catenin, Cy- clinD1, Col I , and Col III were obviously up-regulated in the Ang U group （P 〈0.05）. Compared with the Ang II group, mRNA and protein expressions of Wnt4, Dvl-1, β-catenin, CyclinD1, Col I , and Col III were obviously down-regulated in the middle dose TMP plus Ang II group and the high dose TMP plus Ang II group （P 〈0.05）. The aforesaid indices were dose-dependent in the low, middle, and high dose TMP plus Ang II groups. Conclusion TMP inhibited AngII induced proliferation and collagen secretion of VSMCs through down-regulating Wnt signal pathway.
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2013年第9期1226-1231,共6页
Chinese Journal of Integrated Traditional and Western Medicine
基金
浙江省中医药科学研究基金计划资助项目(No.2011ZA027)
浙江省医药卫生科技计划项目(No.2010KYA155)
关键词
经皮冠状动脉介入
再狭窄
血管平滑肌细胞
增殖
川芎嗪
percutaneous coronary intervention
restenosis
vascular smooth muscle cell
prolifer-ation ：tetramethylpyrazine
