广东医学
廣東醫學
엄동의학
GUNAGDONG MEDICAL JOURNAL
2014年
18期
2814-2816
,共3页
聂成%刘芬%曾振国%詹以安%邓文刚%卿城%邵强%丁成志%揭克敏%钱克俭
聶成%劉芬%曾振國%詹以安%鄧文剛%卿城%邵彊%丁成誌%揭剋敏%錢剋儉
섭성%류분%증진국%첨이안%산문강%경성%소강%정성지%게극민%전극검
肺泡巨噬细胞%miR-146a%核因子-κB%核转位%脂多糖
肺泡巨噬細胞%miR-146a%覈因子-κB%覈轉位%脂多糖
폐포거서세포%miR-146a%핵인자-κB%핵전위%지다당
alveolar macrophage%miRNA-146a%nuclear factor-κB%nuclear translocation%lipopolysaccharide
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。
目的：觀察肺泡巨噬細胞轉染miR-146a後細胞漿與細胞覈內覈因子-κB（ NF-κB）錶達的變化，探討miR-146a調控肺泡巨噬細胞炎癥反應的可能機製。方法將NR8383大鼠肺泡巨噬細胞分成兩組：一組為轉染組，轉染50 nmol／L Pre-miRTM miR-146a Precursors；另一組為對照組，轉染50 nmol CyTM 3-labeled Pre-miRTM Negative Control。兩組細胞轉染24 h後加入含脂多糖培養基（終濃度1μg／mL），培養6 h後收集細胞，分彆提取細胞漿蛋白與細胞覈蛋白，採用Western blot方法檢測NF-κB p65蛋白的錶達。結果轉染組miR-146a錶達較對照組上調（ P＜0.01）。轉染組細胞漿內NF-κB p65蛋白錶達較對照組上調（ P＜0.05），而細胞覈內NF-κB p65蛋白錶達較對照組下調（P＜0.05）。結論肺泡巨噬細胞轉染miR-146a可抑製NF-κB覈轉位，推測miR-146a通過此機製減輕肺泡巨噬細胞炎癥反應。
목적：관찰폐포거서세포전염miR-146a후세포장여세포핵내핵인자-κB（ NF-κB）표체적변화，탐토miR-146a조공폐포거서세포염증반응적가능궤제。방법장NR8383대서폐포거서세포분성량조：일조위전염조，전염50 nmol／L Pre-miRTM miR-146a Precursors；령일조위대조조，전염50 nmol CyTM 3-labeled Pre-miRTM Negative Control。량조세포전염24 h후가입함지다당배양기（종농도1μg／mL），배양6 h후수집세포，분별제취세포장단백여세포핵단백，채용Western blot방법검측NF-κB p65단백적표체。결과전염조miR-146a표체교대조조상조（ P＜0.01）。전염조세포장내NF-κB p65단백표체교대조조상조（ P＜0.05），이세포핵내NF-κB p65단백표체교대조조하조（P＜0.05）。결론폐포거서세포전염miR-146a가억제NF-κB핵전위，추측miR-146a통과차궤제감경폐포거서세포염증반응。
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.
