心脑宁诱导增生VSMC凋亡及对相关基因表达的影响

目的 :探讨活血化瘀中药血清体外诱导增生血管平滑肌细胞凋亡的作用及其分子机制。方法 :采用流式细胞仪分析 ,DNA电泳分析检测细胞凋亡 ;应用Northen印迹检测bcl- 2、c-myc基因表达活性。结果 :经中药血清处理的血管平滑肌细胞 (vascularsmoothmuclecell,VSMC) ,可出现典型的凋亡特征 ,DNA电泳呈梯状图谱 ;流式细胞仪分析显示G0 /G1期细胞阻滞现象 ,亚二倍体凋亡细胞增多 ,出现典型的凋亡峰 ,细胞凋亡率与中药血清浓度具有量效关系。杂交结果提示 ,2 0 %中药血清可抑制抗凋亡基因bcl- 2的表达 ,上调促凋亡基因c -myc的mR NA水平。结论 :活血化瘀中药可通过影响凋亡相关基因bcl- 2、c -myc的表达活性而诱导VSMC凋亡。

Effect of XinNaoNing on Apoptosis and Relevant Gene Expression in Cultivated Vascular Smooth Muscle Cell

Objective :To explore the effect of XinNaoNing recipe on vascular smooth muscle cells(VSMC)apoptosis and its molecular mechanisms. Methods :VSMC apoptosis was detected by flow cytometry and DNA agarose electrophoresis.The expression activity of bcl-2 and c-myc genes were determined using Northern blotting. Results: the typicall characteristics of apoptotic VSMC was observed following treatment by the XinNaoNing drug serum.The DNA extracted from VSMC revealed a ladder pattern in electrophoresis.Flow cytometric analysis revealed G 0/G 1 phenomena,the hypoidploid apoptotic cell increased,displayed typical peak of apoptosis,the rate of cell apoptosis and XinNaoNing serum concentration were in a dose-dependent manner.The result of hybridization showed that 30% of XinNaoNing drug serum could inhibit apoptotic gene bcl-2 expression,upregulated the level of apoptogenous gene c-myc mRNA. Conclusion :VSMC apoptosiss could be induced by XinNaoNing recipe and its mechanism might be through affecting the apoptosiss related gene expression activity.