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2 4h后,免疫印迹法(Westernblot)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2 4h末达到峰值,且T浓度在0 .1～1.0U·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VSMCc fos蛋白的表达;高浓度(1.5×10 - 3mol·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。
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 indicated by Western blot, and TR mRNA expression was evaluated by Reverse Transcription Polymerase Chain Reaction (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 -3 mol·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 -3 mol·L -1 puerarin suppressed this increase. RT-PCR demonstrated that T can significantly increase TR mRNA expression and 1.5 ×10 -3 mol·L -1 puerarin suppressed 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 TR mRNA.
出处
《中国临床药理学与治疗学》
CAS
CSCD
2005年第4期401-406,共6页
Chinese Journal of Clinical Pharmacology and Therapeutics
关键词
葛根素
血管平滑肌细胞
增殖
凝血酶受体
puerarin
vascular smooth muscle cell
proliferation
thrombin receptor
