摘要
目的研究miR-338-3p在内皮细胞EOMA炎症反应中的作用。方法用TNF-α20ng/ml处理内皮细胞系EOMA 24h,实时荧光定量PCR检测miR-338-3p、黏附因子VCAM-1和ICAM-1mRNA表达水平,Western blot分析ERK及p38 MAPK信号通路活性;构建miR-338-3p低表达腺病毒载体(miR-338-3p inhibitor),感染EOMA细胞48h,PCR检测miR-338-3p、VCAM-1及ICAM-1mRNA表达水平,Western blot分析ERK及p38 MAPK信号通路活性。结果 TNF-α20ng/ml处理EOMA细胞24hmiR-338-3p表达水平下降,ERK及p38蛋白磷酸化水平升高,VCAM-1及ICAM-1mRNA表达水平升高;miR-338-3p inhibitor感染EOMA细胞48h,miR-338-3p表达水平明显降低,ERK及p38蛋白磷酸化水平明显升高,黏附因子VCAM-1及ICAM-1mRNA基因表达水平升高。结论在内皮细胞EOMA炎症模型中,TNF-α下调miR-338-3p表达水平;低表达miR-338-3p激活MAPK信号通路活性,增强黏附因子表达水平,诱导EOMA细胞炎症反应。
Objective To study the role of miR-338-3p in the inflammatory response of endothelial cell line EOMA. Methods EOMA cells were treated with TNF-α20 ng/ml,using Real-time PCR to detect the expression levels of miR-338-3p,adhesion factor VCAM-1 and ICAM-1,and western blot analysis of ERK and p38 MAPK signaling pathway ac-tivity.In order to decrease the expression of miR-338-3p,EOMA cells were transfected with miR-338-3p inhibitor for 48 hours.The expression of miR-338-3p and adhesion factor,such as VCAM-1 and ICAM-1 were detected by Real-time PCR.The protein associated with MAPK signaling pathway in this study were analysis of western blot.Results The expression of miR-338-3p was reduced and adhesion factor VCAM-1 and ICAM-1 expression was enhanced by endothe-lial cells EOMA treated with TNF-α20 ng/ml for 24 hours.Western blot analysis of ERK/p38 MAPK signaling path-way activity was enhanced in endothelial cells induced by TNF-α.Down regulation of miR-338-3p expression was ac-companied by enhancing expression of VCAM-1 and ICAM-1 ,and activating MAPK signaling pathway activity in endo-thelial cells EOMA infected with miR-338-3p inhibitor.Conclusion The expression of miR-338-3p was down regulated in EOMA cells induced treated with TNF-α;miR-338-3p inhibitor induced the inflammatory response of endothelial cells by activating the MAPK signaling pathway and enhancing the expression of adhesion molecules.
作者
于淑倩
隋小芳
王凤玲
苑露丹
刘宇
张培培
YU Shu-qian SUI Xiao- Fang WANG Feng-ling et al(Department of Geriatrics, the First Affiliated Hospital of J iamusi University, Jiamusi 154002, China)
出处
《中国实验诊断学》
2016年第12期2002-2005,共4页
Chinese Journal of Laboratory Diagnosis
基金
黑龙江省自然基金面上项目(项目编号:H2015076)
佳木斯大学研究生科技创新重点项目(项目编号:YZ2016_020)
关键词
内皮细胞
炎症反应
黏附因子
miR-338-3p
miR-338-3p
endothelial cells
inflammatory response
adhesion factor
