miRNA-142-3p对肺泡巨噬细胞炎症过程的负向调控及其机制

发布时间：2018-07-01 21:17

本文选题：肺泡巨噬细胞 + 脂多糖类　；参考：《第二军医大学学报》2017年03期

【摘要】：目的探索miRNA-142-3p(miR-142-3p)对脂多糖(LPS)诱导的大鼠肺泡巨噬细胞炎症反应的负向调控作用及其可能机制。方法用100ng/mL LPS诱导肺泡巨噬细胞NR8383,实时定量PCR(qPCR)和蛋白质印迹法分别检测诱导后0、6、24、48h时细胞中miR-142-3p和高迁移率族蛋白(HMGB1)的表达。细胞体外转染miR-142-3p拟似物(miR-142-3p mimic),qPCR检测转染后细胞中miR-142-3p及炎症因子[肿瘤坏死因子α(TNF-α)、白介素(IL)-6、IL-1β和巨噬细胞炎症蛋白2β(MIP-2β)]的表达,蛋白质印迹法检测细胞中HMGB1的表达,酶联免疫吸附(ELISA)法检测细胞培养液中HMGB1的含量。结果LPS诱导NR8383细胞后,细胞中miR-142-3p在48h时表达最高,HMGB1在24h时最高,与0h时相比差异均有统计学意义(P0.05)。过表达miR-142-3p后,NR8383细胞中miR-142-3p的表达升高(P0.05),HMGB1mRNA和蛋白及TNF-α、IL-6、IL-1β和MIP-2βmRNA的表达均降低(P0.05)。结论 miR-142-3p能介导LPS诱导的NR8383细胞的炎症反应过程,该效应可能是通过负向调控HMGB1的表达来实现的。
[Abstract]:Objective to investigate the negative regulation of miRNA-142-3p (miR-142-3p) on the inflammatory response of rat alveolar macrophages induced by lipopolysaccharide (LPS) and its possible mechanism. Methods the expression of miR-142-3p and high mobility group protein (HMGB1) in alveolar macrophages (NR8383) induced by 100 ng / mL LPS was detected by real-time quantitative PCR (qPCR) and Western blotting, respectively. The expression of miR-142-3p and inflammatory cytokines [tumor necrosis factor 伪 (TNF- 伪), interleukin (IL) -6- + IL-1 尾 and macrophage inflammatory protein 2 尾 (MIP-2 尾)] in transfected cells were detected by qPCR, and the expression of HMGB1 was detected by Western blot. Enzyme linked immunosorbent assay (Elisa) was used to detect the content of HMGB1 in cell culture medium. Results after LPS induced NR8383 cells, the expression of miR-142-3p in NR8383 cells was the highest at 48h, which was significantly higher than that at 0h (P0.05). After overexpression of miR-142-3p, the expression of miR-142-3p in NR8383 cells was increased (P0.05), and the expression of HMGB1 mRNA and protein, TNF- 伪 IL-6, IL-1 尾 and MIP-2 尾 mRNA were decreased (P0.05). Conclusion miR-142-3p can mediate the inflammatory response induced by LPS in NR8383 cells, which may be mediated by negative regulation of HMGB1 expression.
【作者单位】：第二军医大学长征医院急救科;
【基金】：国家自然科学基金(81171844)~~
【分类号】：R563

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