丹酚酸B对MC3T3-E1细胞Cbfa1基因表达的影响及Cbfa1启动子活性的调节.docVIP

 丹酚酸 B 对 MC3T3-E1 细胞 Cbfa1 基因表 达的影响及 Cbfa1 启动子活性的调节# 5 10 15 20 25 30 35 40  摘要：目的 探讨丹酚酸 B 对小鼠 MC3T3-E1 细胞核结合因子 a1（Cbfa1）基因 mRNA 表达 和 Cbfa1 启动子活性的影响。方法 分别采用不同浓度丹酚酸 B 干预小鼠 MC3T3-E1 前成骨 细胞系 24 和 48 小时，real-time PCR 检测丹酚酸 B 对细胞中 Cbfa1 基因 mRNA 表达的影响； 构建小鼠 Cbfa1 基因启动子荧光素酶报告基因载体并转染 MC3T3-E1 细胞，通过检测荧光素 酶活性观察丹酚酸 B 对 Cbfa1 启动子活性的调节。结果 1μM 和 10μM 的丹酚酸 B 分别处理 MC3T3-E1 细胞 24 小时和 48 小时，可上调 Cbfa1 mRNA 的表达（F=23.24，P0.01），其 中以 1μM 丹酚酸 B 处理 48 小时上调效应最显著，约为对照组的 1.71 倍（t=10.36，P＜0.01）。 1μM 和 10μM 的丹酚酸 B 干预 MC3T3-E1 细胞后，Cbfa1 启动子活性升高，分别为对照组的 2.63、1.89 倍（F=13.27，t 分别为 5.83、4.53，P0.01）。结论 丹酚酸 B 可促进 MC3T3-E1 细胞中 Cbfa1 基因表达并上调 Cbfa1 启动子活性。 关键词：中西医结合医学；丹酚酸 B；核结合因子 a1；基因表达；启动子 中图分类号：R34 Action of Salvianolic Acid B on Cbfa1 Gene Expression in MC3T3-E1 Cells and Activity of Cbfa1 Promoter ZHENG Fang1, WANG Hongwu1, BAO Liying1, ZHAO Shuwu1, Pang Xiaoli2 (1. Traditional Chinese Medicine School,Tianjin University of Tradtional Chinese Medicine, TianJin 300193; 2. Nursing School, Tianjin University of Tradtional Chinese Medicine, TianJin 300193) Abstract: Objective Explore the effects of salvianolic acid B(SalB) on the mRNA expression of Cbfa1 gene in MC3T3-E1 cell line and the activity of Cbfa1 promoter. Methods The mouse MC3T3-E1 cells were treated for 24hours or 48hours with SalB of different concentration respectively. The mRNA expression of Cbfa1 gene were examined by using real-time RT-PCR. Luciferase reporter vector of mice Cbfa1 promoter was constructed and then transfected into MC3T3-E1 cells. The action of SalB on the activity of Cbfa1 promoter were evaluated by monitoring the expression of luciferase. Results We demonstrated in MC3T3-E1cells, 0.1μM and 10μM SalB increased Cbfa1 mRNA expression after treatment for 24hours and 48hours（F=23.24， P0.01）. The maximum increase of Cbfa1mRNA level is 1.71-fold greater than that of contol under intervention of 0.1μM SalB for 48hours（t=10.36，P＜0.01）. Compared with the control, the promoter activity of Cbfa1 gene were upregulated to 2.63-fol