普罗布考抑制大鼠血管平滑肌细胞周期素D1蛋白表达和G1→S转换  

Probucol Reduces Cyclin D1 Protein Expression and G1→S Transition in Rat Vascular Smooth Muscle Cells

作　　者：涂永生[1] 严鹏科[1] 朱炳阳[1] 黄红林[1] 廖端芳[1] 

机构地区：[1]南华大学药物药理研究所

出　　处：《中国动脉硬化杂志》2005年第6期676-680,共5页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金(30171084)资助;国家973子项目(G2000056905)资助

摘　　要：目的研究普罗布考是否通过阻滞细胞周期抑制平滑肌细胞增殖.方法首先用普罗布考和氧化型低密度脂蛋白与平滑肌细胞共同孵育.然后用MTT法和细胞计数法测定细胞增殖,用流式细胞术观察细胞周期,最后用蛋白印迹测蛋白表达.结果 MTT法和细胞计数法显示25～100 mg/L氧化型低密度脂蛋白都能刺激平滑肌细胞增殖,效应呈浓度依赖性,普罗布考能显著抑制氧化型低密度脂蛋白所诱导的细胞增殖,效应呈浓度和时间依赖性.流式细胞术分析表明普罗布考增加了G0/G1 细胞比例达28.6% (P<0.01,n=3),减少S期细胞比例达83.5% (P<0.01, n=3).蛋白印迹结果显示氧化型低密度脂蛋白诱导细胞周期素D1蛋白表达,高峰在6～12 h,普罗布考显著减少周期素D1蛋白表达,在6 h和12 h分别减少达26% 和23% (P均<0.01,n=3).结论普罗布考显著抑制氧化型低密度脂蛋白所诱导的细胞增殖,其抗增殖活性与普罗布考下调周期素D1蛋白表达,从而阻滞细胞由G0/G1 期向S期转换有关.Aim To examine whether procucol could inhibit vascular smooth muscle ceils （VSMC） proliferation by impairing cell cycle progression. Methods Initially, quiescent rat VSMC were treated with probucol in the presence of oxidized low density lipoprotein （ox-LDL）. Then cell proliferation was measured by MTT assay and cell counts, and cell cycle distribution was analyzed by Flow-cytometry （ FCM）. Finally, protein expression was examined by western blot. Results MTT assay and cell counts revealed that 25 mg/L,50 mg/L and 100 mg/L ox-LDL stimulated VSMC proliferation in a dose dependent manner. Probucol significantly inhibited VSMC proliferation induced by ox-LDL in a dose and time dependent manner. FCM analysis showed that probucol increased cells in C0/G1 by 28.6% （ P 〈 0.01, n = 3）, reduced the cells in S by 83.5% （ P 〈 0.01, n = 3 ）. Also, 100 mg/L ox-LDL induced cyclin D1 expression, with peak in 6- 12 h. Probucol markedly inhibited cyclin DI expression by 26% at6 h and 23% at 12 h respectively （both P〈0.01,n=3）. Conlusions Probucol significandy inhibits ox-LDL induced VSMC proliferation. Tiffs antiprolfferative effect of probucol is correlated with suppression of cell progression by blocking the cell cycle in C0/G1 . Furthermore, suppression of cell cycle progression is associated with the down-expression of cyclin D1 protein by probucol

关 键 词：药理学 普罗布考 平滑肌细胞 细胞周期 细胞增殖 

分 类 号：R972.6[医药卫生—药品]