miRNA-17-5p在免疫细胞炎症反应和自噬中的作用

发布时间：2018-04-24 23:30

本文选题：脂多糖 + 炎症　；参考：《广东医学》2016年12期

【摘要】：目的探讨脂多糖(LPS)诱导免疫细胞炎症反应和自噬中miRNA-17-5p的调控作用。方法体外培养人白血病单核细胞株(THP-1),100 ng/m L佛波醇将THP-1细胞诱导24 h分化成为巨噬细胞,设置空白对照组和LPS刺激组,其中LPS刺激组加500 ng/m L的LPS,空白对照组只加灭菌水。刺激THP-1巨噬细胞12 h后采用酶联免疫吸附法(ELISA)检测细胞培养上清液中肿瘤坏死因子-α(TNF-α)、单核细胞趋化蛋白-1(MCP-1)蛋白水平;采用定量PCR反应检测基因miRNA-17-5p、PTEN的表达情况,采用Western blot检测P62蛋白水平。结果与空白对照组比较,LPS刺激组培养上清液中TNF-α、MCP-1蛋白分泌量显著增加(P0.01),PTEN表达显著下调(P0.01),细胞内miRNA-17-5p显著上调(P0.01),细胞内P62蛋白表达上调(P0.05)。结论 miRNA-17-5p参与LPS诱导的免疫细胞炎症反应和自噬调控。
[Abstract]:Objective to investigate the inflammatory response of immune cells induced by lipopolysaccharide (LPS) and the regulation of miRNA-17-5p in autophagy. Methods Human leukemic monocytes were cultured in vitro. THP-1 cells were induced to differentiate into macrophages for 24 h by 100 ng/m / L phorbol alcohol. The blank control group and LPS stimulated group were divided into two groups: LPS stimulated group plus 500 ng/m L LPSs, and blank control group only added sterilizing water. After 12 h stimulation of THP-1 macrophages, the levels of tumor necrosis factor- 伪 (TNF- 伪) and monocyte chemoattractant protein (MCP-1) in the supernatant of cell culture were detected by Elisa, and the expression of miRNA-17-5pPTEN gene was detected by quantitative PCR reaction. The level of P62 protein was detected by Western blot. Results compared with the control group, the secretion of TNF- 伪 MCP-1 protein increased significantly in the culture supernatant of LPS-stimulated group, the expression of P0.01PTEN was down-regulated, the expression of miRNA-17-5p was up-regulated by P0.01T, and the expression of P62 protein was upregulated by P0.05A. Conclusion miRNA-17-5p is involved in LPS induced immune cell inflammation and autophagy.
【作者单位】：广州医科大学附属第二医院健康体检中心;广东省妇幼保健院妇产科;广州医科大学附属第二医院重症医学科;
【基金】：广东省科技计划项目(编号:2012B031800233)
【分类号】：R363

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