MicroRNA-132转染对脂多糖诱导的肺泡巨噬细胞炎症反应的作用

 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。

Effect of microRNA-132 transfection on lipopolysaccharide-induced inflammation in rat alveolar macrophages

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 without pyrogen in vitrowere 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 respectively. The cell proliferation was detected by Cell Counting Kit-8 (CCK-8) assay. Real-time PCR was used to detect the expression 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 interleukin-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 transfected 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-α and IL-6 in transfected NR8383 cells were decreased compared with blank control group and negative control group (P<0.05).CONCLUSION: Transfection of alveolar macrophages with miR-132 significantly suppresses the cell growth, and inhibits inflammatory responses induced by LPS.