转染miR-146a对肺泡巨噬细胞中核因子-κB表达的影响

摘要

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.%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-miRTM miR-146a Precursors；另一组为对照组，转染50 nmol CyTM 3-labeled Pre-miRTM Negative 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。