MicroRNA-132转染对脂多糖诱导的肺泡巨噬细胞炎症反应的作用

摘要

[ ABSTRACT] AIM:To investigate the effect of microRNA-132 ( miR-132 ) transfection on the lipopolysaccharide ( LPS)-induced inflammation in rat alveolar macrophages.METHODS:The rat alveolar macrophage NR8383 cultured with-out pyrogen in vitro were divided into blank control group, negative control group and transfected group.The cells in the 3 groups were transfected with phosphate buffer solution ( PBS) , Lipofectamine 2000 and synthesized miR-132 mimic respec-tively.The cell proliferation was detected by Cell Counting Kit-8 ( CCK-8) assay.Real-time PCR was used to detect the ex-pression of miR-132 in the cells.After NR8383 cells were stimulated with LPS for 6 h, the NF-κB DNA-binding activity was measured by electrophoretic mobility shift assay ( EMSA) .The expression of tumor necrosis factor-α( TNF-α) and interleu-kin-6 (IL-6) in NR8383 cells was assayed by Western blotting.RESULTS: After transfection, the expression of miR-132 was significantly higher than that in blank control group and negative control group.The growth of NR8383 cells in transfect-ed group was significantly inhibited compared with blank control group and negative control group ( P<0.05 ) .After the cells were stimulated with LPS, the productions of NF-κB, TNF-αand IL-6 in transfected NR8383 cells were decreased com-pared with blank control group and negative control group (P<0.05).CONCLUSION:Transfection of alveolar macropha-ges with miR-132 significantly suppresses the cell growth, and inhibits inflammatory responses induced by LPS.%目的：探讨转染微小RNA-132（miR-132）对肺泡巨噬细胞炎症反应的作用。方法：将体外去致热源培养的大鼠肺泡巨噬细胞株NR8383分为空白对照组、阴性对照组和转染组，分别采用miR-132增敏剂、错义链和PBS作用。转染24 h后，CCK-8法检测细胞増殖；实时荧光定量PCR检测细胞中miR-132的表达；用脂多糖（ LPS）作用细胞后，凝胶电泳迁移率实验（ EMSA ）检测细胞中NF-κB活性；Western blotting法检测细胞中肿瘤坏死因子α（TNF-α）和白细胞介素6（IL-6）的表达。结果：与空白对照组和阴性对照组相比较，转染组细胞中miR-132的表达明显升高；转染组细胞増殖被明显抑制，与空白对照组相比较，差异有统计学意义（ P＜0.05）；LPS作用后，转染组NF-κB、TNF-α和IL-6表达量显著下降，与空白对照组和阴性对照组相比较，差异均有统计学意义（ P ＜0.05）。结论：转染miR-132可抑制NR8383细胞増殖，并抑制LPS诱导的NR8383细胞的炎症反应。