miR-27b对IL-17诱导下MCP-1表达的影响作用  

Effect of miR-27b on the expression of MCP-1 induced by IL-17

作　　者：卫肖[1] 沈燕[1] 张凤岐[1] 

机构地区：[1]郑州大学第一附属医院检验科,郑州450052

出　　处：《现代免疫学》2014年第2期146-150,共5页Current Immunology

摘　　要：微小RNA(microRNA,miRNA)是一种基因表达调控因子,本研究旨在研究miR-27b对IL-17诱导下的单核细胞趋化蛋白-1(MCP-1)表达的影响。我们转染miR-27bmimic于H9C2细胞中,采用SYBR green I荧光定量PCR检测H9C2心肌细胞中miR-27b和MCP-1基因的表达情况,以ELISA分析H9C2细胞培养上清液中MCP-1蛋白的表达。在IL-17对MCP-1表达的影响的研究中,结果显示,IL-17作用2h后MCP-1mRNA的表达量开始升高,4h时达到高峰,尔后开始下降,而MCP-1蛋白的表达量0~24h逐渐升高。在miR-27b对IL-17诱导下MCP-1表达的影响的研究中,结果显示,IL-17组与空白对照组比较,MCP-1mRNA和蛋白水平显著升高(P<0.05);IL-17组与转染miRNA阴性对照+IL-17组比较,两组的MCP-1mRNA和蛋白水平无显著差异(P>0.05);转染miR-27bmimic+IL-17与转染miRNA阴性对照+IL-17组比较,MCP-1mRNA和蛋白水平显著下降(P<0.01)。本研究表明,(1)IL-17可上调MCP-1的表达,这与IL-17的作用时间有关。(2)过表达miR-27b可下调IL-17诱导下MCP-1的表达水平。Small RNA （MicroRNA, miRNA） is an element that regulates gene expression. In this study we further investiga- ted the effect of miR-27b on the expression of monocyte chemoattractant protein -1 （MCP-1） induced by IL-17. We transfected miRNA oligo into H9C2 cardiomyocytes and the expression of miR-27b and MCP-1 mRNA in H9C2 cardiomyocytes were de- tected by SYBR green I fluorescence quantitative PCR. Enzyme linked immunosorbent assay （ELISA） was used to measure the concentration of MCP-1 protein in the culture supernatant of H9C2 cells. In terms of the effect of IL-17 on the expression of MCP-1, the results showed that MCP-1 mRNA expression began to increase at 2h after stimulation by IL-17 and reached a peak at 4h, and thereafter began to decline. The amount of MCP-1 protein increased gradually in 0N 24 h. In the research of the effect of miR-27b on the expression of MCP-1 induced by IL-17, the results showed that MCP-1 mRNA and protein levels were significantly increased in H9C2 cells stimulated by IL-17 as compared with the control group（P^0.05）~ while there were no differences in the expression of MCP-1 mRNA and protein in IL-17 group as compared with transfected with miRNA negative control and IL-17 group （P~ 0.05）, the cells transfected with miR-27b mimic and IL-17 compared with that of miRNA nega- tive control and IL-17 group, MCP-1 mRNA and protein levels were significantly decreased （P%0.01）. This study demon- strates that IL-17 could up-regulate the expression of MCP-1 in a time dependent manner and over-expression of miR-27b might negatively regulate the expression of MCP-1 induced by IL-17.

关 键 词：H9C2心肌细胞 miR-27b MCP-1 IL-17 

分 类 号：R392.11[医药卫生—免疫学]