转染miR-146a对肺泡巨噬细胞中核因子-κB表达的影响

Effects of miR-146a transfection on nuclear factor-κB expression of alveolar macrophage

目的观察肺泡巨噬细胞转染miR-146a后细胞浆与细胞核内核因子-κB(NF-κB)表达的变化,探讨miR-146a调控肺泡巨噬细胞炎症反应的可能机制。方法将NR8383大鼠肺泡巨噬细胞分成两组:一组为转染组,转染50 nmol/L Pre-miRTMmiR-146a Precursors;另一组为对照组,转染50 nmol CyTM3-labeled PremiRTMNegative Control。两组细胞转染24 h后加入含脂多糖培养基(终浓度1μg/mL),培养6 h后收集细胞,分别提取细胞浆蛋白与细胞核蛋白,采用Western blot方法检测NF-κB p65蛋白的表达。结果转染组miR-146a表达较对照组上调(P<0.01)。转染组细胞浆内NF-κB p65蛋白表达较对照组上调(P<0.05),而细胞核内NF-κB p65蛋白表达较对照组下调(P<0.05)。结论肺泡巨噬细胞转染miR-146a可抑制NF-κB核转位,推测miR-146a通过此机制减轻肺泡巨噬细胞炎症反应。

Objective To explore the plausible mechanism that miR-146a modulates the inflammatory response in alveolar macrophage by comparing nuclear factor-κB （ NF-κB） expression between cytoplasm and nucleus.Methods Alveolar macrophages from rat （NR8383） were divided into two groups： Group Transfected （transfected with Pre -miRTM miR -146a precursors） and Group Control （transfected with CyTM3 -labeled Pre -miRTM negative control）.After24 -hour transfection, both groups were cultured in complete medium with lipopolysaccharide charge for 6 hours.NF -κBp65 proteins from cytoplasm and nucleus were extracted and determined by Western blot.Results Compared to CroupControl, miR -146a expression of Group Transfected was significantly up -regulated （P 〈0.01）, so was the Compared toCroup Control, the cytoplasm NF -κB p65 expression of Group Transfected was also significantly up -regulated （P 〈0.05）, while the nuclear NF -κB p65 expression was significantly down -regulated （P 〈0.05）.Conclusion Transfectionof miR -146a into alveolar macrophage inhibits the NF -κB′s nuclear translocation, suggesting a mechanism viawhich miR -146a alleviates the inflammatory response in alveolar macrophages.