摘要
目的 :探讨益气健脾中药血清体外诱导血管平滑肌细胞凋亡的作用及其分子机制。方法 :采用缺口末端标记 (terminaldeoxynucleotidyltransferasemediateddUTPnickedlabeling ,TUNEL)分析、透射电镜、流式细胞仪分析、DNA电泳分析检测细胞凋亡 ;应用Northern印迹检测bcl 2、c myc和 p5 3基因表达活性。结果 :经中药血清处理的血管平滑肌细胞 (vascularsmoothmusclecell,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 ,30 %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和 p5 3mRNA水平。 结论 :益气健脾中药通过影响凋亡相关基因bcl 2、c myc和p5
Objective: To explore the effect of Yiqi Jianpi (YQJP, supplementing Qi and invigorating Spleen) recipe on vascular smooth muscle cells (VSMC) apoptosis and it′s mechanisms. Methods: VSMC apoptosis was detected by terminal deoxynucleotidyl transferase mediated dUTP nicked labeling (TUNEL) analysis, transmission electron microscope (TEM), flow cytometry and DNA agarose electrophoresis. The expression activity of bcl 2, c myc and p53 genes were determined using Northern blotting. Results: The typical characteristics of apoptotic VSMC was observed following treatment by the YQJP drug serum. The DNA extracted from VSMC revealed a ladder pattern in electrophoresis. Flow cytometric analysis revealed G 0/G 1 stage cell blocking phenomena, the hypodiploid apoptotic cell increased, displayed typical peak of apoptosis, the rate of cell apoptosis and YQJP serum concentration were in a dose dependent manner. The result of hybridization showed that 30% of YQJP serum could inhibit apoptotic gene bcl 2 expression, upregulated the level of apoptogenous gene c myc and p53 mRNA. Conclusion: VSMC apoptosis could be induced by YQJP recipe and its mechanism might be through affecting the apoptosis related gene expression activity.
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2001年第12期909-912,共4页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家科技部攀登特别支持项目 (国科基字 [1 999] 0 4 5号 )
国家自然科学基金项目 (No .39970 2 74)
河北省自然科学基金项目 (No.30 1 583)
关键词
益气健脾
血清药理学
血管平滑肌细胞
凋亡
supplementing Qi and invigorating Spleen, seropharmacology, vascular smooth muscle cells, apoptosis
