丹参酮Ⅱ_A抑制大鼠血管平滑肌细胞增殖及其机制研究  

Inhibitory effects and mechanism of tanshinone Ⅱ_A on proliferation of rat aortic smooth muscle cells

作　　者：李欣[1] 杜俊蓉[2] 白波[2] 余彦[2] 郑晓媛[2] 杨芳[2] 郑虎[2] 

机构地区：[1]天津医科大学基础医学院,天津300070 [2]四川大学华西药学院,四川成都610041

出　　处：《中国中药杂志》2008年第17期2146-2150,共5页China Journal of Chinese Materia Medica

摘　　要：目的:观察丹参酮ⅡA(tanshinoneⅡA,TA)对10%胎牛血清诱导的大鼠血管平滑肌细胞(RVSMC)增殖的抑制作用,并探讨其可能的作用机制。方法:体外培养大鼠主动脉平滑肌细胞A7r5细胞株,以终浓度为10%的胎牛血清(FBS)作为刺激因素,用细胞计数法、噻唑蓝(MTT)比色法和5-溴脱氧尿嘧啶(BrdU)掺入法测定TA对细胞增殖的影响,用流式细胞术(FCM)分析细胞周期分布特征,用Western blot实验测定细胞外信号调节激酶1/2(ERK1/2)磷酸化活性,用RT-PCR测定c-fos的表达水平。结果:细胞计数、MTT比色法和BrdU掺入实验表明丹参酮ⅡA能抑制10%FBS所诱导的VSMC增殖,作用强度呈剂量依赖性;细胞周期分析显示,TA处理组G0/G1期细胞百分比高于10%FBS组,而S期比例低于10%FBS组,表明TA可阻止10%FBS所诱导的细胞周期由G0/G1期向S期推进;Western blot结果显示与10%FBS组相比,TA处理组ERK1/2磷酸化活性降低;RT-PCR结果显示TA处理组c-fos表达水平降低。结论:丹参酮可抑制10%FBS诱导的体外培养大鼠动脉平滑肌细胞增殖,此作用可能与其阻止细胞周期由G0/G1期向S期推进,抑制MAPK信号转导通路激活,进而下调c-fos表达有关。Objective： To investigate the effects of tanshinone Ⅱ A （ TA, one of the active components of Salvia miltiorrhiza Bge）, on the proliferation of cultured rat vascular smooth muscle cells （VSMC）, and to clarity the probable mechanism. Method： Cell culture technique was used in vitro and treated with or without 10% FBS. The inhibitory effect of TA on proliferation of VSMC was observed by cell count, MTT metabolism measuring and BrdU incorporation assay. Flow cytometry was performed to track cell cycle progression. Western bolts were performed to evaluate ERK1/2 activity. The expression of c-fos was examined by RT-PCR. Result： The results of cell number, MTT assay and BrdU incorporation showed that TA cound significantly inhibit 10% FBS induced proliferation in a dose-dependent manner. Flow cytometry （FCM） analysis indicated that the G0/G1 phase fraction ratio of TA group was higher than that of 10% FBS group, while the S-phase fraction ratio was lower than that of 10% FBS group. Western blot＇s results displayed that TA inhibited the phosphorylation of ERK1/2, and in accordance with this findings. TA could decrease the early elevation of c-fos expression. Conclusion： These results suggest that TA can inhibit 10% FBS induced the proliferation of VSMC. This effect may be related to its blocking VSMCs cell cycle in G0/G1 phase, inhibiting of the ERK1/2 activity, and decreasing the expression of c-fos.

关 键 词：丹参酮ⅡA 平滑肌细胞 细胞周期 丝裂素活化蛋白激酶 c—los 

分 类 号：R285.5[医药卫生—中药学]