摘要
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。
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 Ceil 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 LIPS for 6 h, the NF-KB DNA-binding activity was measured by electrophoretic mobility shift assay （EMSA）. The expression of tumor necrosis factor-or （TNF-ct） 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-KB, TNF-ot and 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.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2014年第12期2190-2194,共5页
Chinese Journal of Pathophysiology
基金
浙江省卫生厅资助项目(No.2012A122)
