葛根素对血管平滑肌细胞增殖及对 c-fos 蛋白和凝血酶受体 mRNA 表达的影响

中国临床药理学与治疗学 ›› 2005, Vol. 10 ›› Issue (4): 401-406.

葛根素对血管平滑肌细胞增殖及对 c-fos 蛋白和凝血酶受体 mRNA 表达的影响

许轶洲, 李佩璋, 王宁夫, 徐海鹰, 彭文辉

杭州市第一人民医院心血管内科, 杭州 310006, 浙江

收稿日期:2004-12-02 修回日期:2005-03-29 出版日期:2005-04-26 发布日期:2020-11-19
通讯作者: 许轶洲,女, 硕士, 在读博士, 主治医师, 主要从事心血管病学及血管性疾病基础研究。E-mail:13306811689@zj165.com

Effects of puerarin on proliferation of vascular smooth muscle cells and c-fos protein expression and TR mRNA expression

XU Yi-zhou, LI Pei-zhang, WANG Ning-fu, XU Hai-ying, PENG Wen-hui

Department of Cardiology, the First People's Hospital of Hangzhou, Hangzhou310006, Zhejiang, China

Received:2004-12-02 Revised:2005-03-29 Online:2005-04-26 Published:2020-11-19

摘要/Abstract

摘要： 目的: 通过建立凝血酶(T) 诱导的血管平滑肌细胞(VSMC) 增殖模型, 观察葛根素对 T 诱导的VSMC 增殖的影响, 并进一步观察其对c-fos 蛋白以及凝血酶受体(TR) mRNA 表达的作用, 旨在认识葛根素作用的分子机制。方法: 以细胞计数法和流式细胞仪测定 DNA 含量, 细胞周期分析观察T 及葛根素对VSMC增殖和DNA 合成的影响。T 及葛根素等各处理因素作用 24 h 后, 免疫印迹法(Western blot)检测 c-fos 蛋白表达, 半定量逆转录聚合酶链式反应(RT-PCR) 检测 TRmRNA 的表达。结果: T 对 VSMC有明显促增殖作用, 促增殖效应在 24 h 末达到峰值, 且 T 浓度在 0.1 ~ 1.0 U°L^-1 有剂量依赖关系;葛根素呈剂量依赖性地抑制 T 诱导的细胞增殖、DNA 合成以及 VSMC c-fos 蛋白的表达;高浓度(1.5×10^-3 mol°L^-1) 的葛根素可显著抑制 T 诱导的TRmRNA 上调。结论: 葛根素能抑制 T 诱导的VSMC增殖, 这可能与其抑制 c-fos 蛋白有关, 并部分与其抑制 TRmRNA 表达有关。

关键词: 葛根素, 血管平滑肌细胞, 增殖, 凝血酶受体

Abstract: AIM: To observe the effects of puerarin on the proliferation of vascular smooth muscle cells (VSMC) induced by thrombin and the effects of puerarin on the c-fos protein expression and thrombin receptor (TR) mRNA expression.METHODS: DNA contents were detected by cell number and cell cycle analysis and effects of puerarin on proliferation of VSMC were detected by flow cytometry. The changes of c-fos protein was indi-cated by Western blot, and TR mRNA expression was e-valuated by Reverse Transcription Polymerase Chain Re-action (RT-PCR) which was treated with T and puerarin after 24 hours. RESULTS: T increased the cell numbers of VSMC, the peak of proliferation curve was at 24 h, and the effects of T was in a dose-dependent manner (0.1 -1.0 U°L^-1). 1.5 ×10^-5 -1. 5 ×10^-3mol°L^-1 puerarin significantly suppressed this stimulation of VSMC proliferation and DNA synthesis induced by T. Western blot demonstrated that T increased c-fos protein which treated with T and puerarin after 24 hour. 1.5 ×10^-5 -1.5 ×10^-3mol·L^-1 puerarin suppressed thisincrease. RT-PCR demonstrated that T can significantly increase TR mRNA expression and 1.5×10^-3mol°L^-1 puerarin sup-pressed this increase. CONCLUSION: Puerarin can suppress the proliferation and DNA synthesis of VSMC promoted by T. The inhibitory effects of puerarin are closely related with the suppression of c-fos protein, and partly related with the suppression on TRmRNA.

Key words: puerarin, vascular smooth muscle cell, proliferation, thrombin receptor

中图分类号:

R285

许轶洲, 李佩璋, 王宁夫, 徐海鹰, 彭文辉. 葛根素对血管平滑肌细胞增殖及对 c-fos 蛋白和凝血酶受体 mRNA 表达的影响[J]. 中国临床药理学与治疗学, 2005, 10(4): 401-406.

XU Yi-zhou, LI Pei-zhang, WANG Ning-fu, XU Hai-ying, PENG Wen-hui. Effects of puerarin on proliferation of vascular smooth muscle cells and c-fos protein expression and TR mRNA expression[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2005, 10(4): 401-406.

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