NF-κB信号通路阻断对内皮细胞增殖的抑制作用观察  

Inhibition effect of knockdown of NF-κB signaling pathway by small interference RNA on endothelial cell proliferation

作　　者：何晓鹏[1] 毕研文[2] 刘相燕[1] 王洲 王静[1] 梁雪红[1] 刘凡英[1] 

机构地区：[1]山东大学附属省立医院,济南250021 [2]山东大学齐鲁医院

出　　处：《山东医药》2011年第38期25-27,共3页Shandong Medical Journal

摘　　要：目的观察内皮细胞的核转录因子(NF)-κB信号通路阻断对内皮细胞增殖的抑制作用。方法构建NF-κB p65的特异性小分子干扰RNA(siRNA)真核表达载体。取体外培养的大鼠主动脉内皮细胞(SVAREC细胞),随机分为空白对照组、脂多糖(LPS)组、LPS+HK组和LPS+p65 siRNA组。除空白对照组外,均予1μg/mlLPS诱导孵育。LPS+HK组和LPS+p65 siRNA组分别转染无义序列HK和p65 siRNA真核表达载体。继续培养72 h后用Western blot法检测各组细胞p65蛋白,流式细胞术测算细胞周期,MTT法测算内皮细胞增殖指数及细胞增殖率。结果 NF-κB p65蛋白表达水平量LPS组为0.913±0.073,较空白对照组0.527±0.053明显增高(P<0.05);LPS+HK组为0.926±0.065,与LPS组相比P>0.05,LPS+p65 siRNA组为0.473±0.071,与LPS组相比P<0.05。LPS组G0/G1期细胞比例较空白对照组显著降低、S期细胞比例、细胞增殖指数及细胞增值率较对照组显著增高(P均<0.05);LPS+p65 siRNA组细胞与LPS组相比G0/G1期比例显著增高,S期细胞比例、细胞增殖指数及细胞增殖率显著降低(P均<0.05),而LPS+HK组与LPS组比较P均>0.05。结论 NF-κB信号传导通路阻断可抑制内皮细胞增殖。Objective To observe the inhibition effect of small interfering RNA（siRNA） targeting nuclear factor-κB（NF-κB） on endothelial cell proliferation according to its downregulation of NF-κB pathway.Methods The siRNA eukaryotic expression vector targeting rat NF-κB p65 was constructed and transfected into the cultured rat endothelial cells.After transfection,cells were divided into different groups and cultured with or without LPS.Following that,the p65 protein expression in the cytoplasma was detected by Western blotting.Furthermore,the proliferative rate of the cell growth was detected by MTT assay and the distribution of the cell cycle was detected by flow cytometry.Results The p65 siRNA effectively downregulated the protein level of p65（P0.05）.Meanwhile,the proliferation of the cells transfected with p65 siRNA expression vector was significantly inhibited（P0.05）,and the ratio of cells at G0/G1 stage was markedly increased,while the ratio at S stage was significantly decreased in the transfected cells as compared with that in the control（P0.05）.Conclusion Knockdown of NF-κB signaling pathway by small interference RNA can inhibit endothelial cell proliferation.

关 键 词：核转录因子-ΚB RNA干扰 小分子干扰RNA 内皮细胞 细胞增殖 

分 类 号：R654.3[医药卫生—外科学]