中国实用医刊
中國實用醫刊
중국실용의간
CENTRAL PLAINS MEDICAL JOURNAL
2010年
11期
1-3,7
,共4页
骨髓间质干细胞%神经元样细胞%小凹蛋白-1%RNA干扰
骨髓間質榦細胞%神經元樣細胞%小凹蛋白-1%RNA榦擾
골수간질간세포%신경원양세포%소요단백-1%RNA간우
Marrow mesenchymal stem cells%Neuron-like cells%Caveolin-1%RNA interfering
目的 应用RNA干扰(RNA interference)技术,抑制沉默小凹蛋白-1(caveolin-1)基因的表达,观察小凹蛋白-1在骨髓间充质干细胞(MSCs)分化为神经元样细胞的作用.方法 构建大鼠caveolin-1 siRNA(small interfering RNA),然后传染大鼠MSCs,诱导分化并在倒置显微镜下进行形态学观察,用MTT比色法检测细胞存活率,采用免疫细胞化学染色法检测Nestin、 NSE(神经元标记物) 、GFAP(神经胶质细胞标记物)的表达,设诱导未传染组和传染对照组作为对照研究.结果 用caveolin-1 siRNA传染MSCs后,caveolin-1基因表达消失,传染后24 h和3 d MSCs细胞存活率下降,与诱导未传染组和传染对照组比较差异有统计学意义(P<0.01),传染caveolin-1 siRNA的MSCs向神经元样细胞分化效率显著提高,诱导24 h后NSE阳性细胞率为(75.5±2.5)%,6 d后达(81.6±3.5)%,且未见GFAP的表达,与诱导未传染组和传染对照组比较差异有统计学意义(P<0.01).结论 caveolin-1 siRNA抑制caveolin-1基因的表达,可提高MSCs向神经元样细胞的分化效率.
目的 應用RNA榦擾(RNA interference)技術,抑製沉默小凹蛋白-1(caveolin-1)基因的錶達,觀察小凹蛋白-1在骨髓間充質榦細胞(MSCs)分化為神經元樣細胞的作用.方法 構建大鼠caveolin-1 siRNA(small interfering RNA),然後傳染大鼠MSCs,誘導分化併在倒置顯微鏡下進行形態學觀察,用MTT比色法檢測細胞存活率,採用免疫細胞化學染色法檢測Nestin、 NSE(神經元標記物) 、GFAP(神經膠質細胞標記物)的錶達,設誘導未傳染組和傳染對照組作為對照研究.結果 用caveolin-1 siRNA傳染MSCs後,caveolin-1基因錶達消失,傳染後24 h和3 d MSCs細胞存活率下降,與誘導未傳染組和傳染對照組比較差異有統計學意義(P<0.01),傳染caveolin-1 siRNA的MSCs嚮神經元樣細胞分化效率顯著提高,誘導24 h後NSE暘性細胞率為(75.5±2.5)%,6 d後達(81.6±3.5)%,且未見GFAP的錶達,與誘導未傳染組和傳染對照組比較差異有統計學意義(P<0.01).結論 caveolin-1 siRNA抑製caveolin-1基因的錶達,可提高MSCs嚮神經元樣細胞的分化效率.
목적 응용RNA간우(RNA interference)기술,억제침묵소요단백-1(caveolin-1)기인적표체,관찰소요단백-1재골수간충질간세포(MSCs)분화위신경원양세포적작용.방법 구건대서caveolin-1 siRNA(small interfering RNA),연후전염대서MSCs,유도분화병재도치현미경하진행형태학관찰,용MTT비색법검측세포존활솔,채용면역세포화학염색법검측Nestin、 NSE(신경원표기물) 、GFAP(신경효질세포표기물)적표체,설유도미전염조화전염대조조작위대조연구.결과 용caveolin-1 siRNA전염MSCs후,caveolin-1기인표체소실,전염후24 h화3 d MSCs세포존활솔하강,여유도미전염조화전염대조조비교차이유통계학의의(P<0.01),전염caveolin-1 siRNA적MSCs향신경원양세포분화효솔현저제고,유도24 h후NSE양성세포솔위(75.5±2.5)%,6 d후체(81.6±3.5)%,차미견GFAP적표체,여유도미전염조화전염대조조비교차이유통계학의의(P<0.01).결론 caveolin-1 siRNA억제caveolin-1기인적표체,가제고MSCs향신경원양세포적분화효솔.
Objective To investigate the role of caveolin-1 in bone marrow mesenchymal stem cells(MSCs) differentiating into neuron-like cells by transfecting MSCs with small interfering RNA(siRNA). Methods Construct caveolin-1 siRNA and then transfect into MSCs cultured in vitro. Cell growth was observed by an inverted phase contrast microscope. The survival ratio of MSCs was determined by MTT before transfection,24 hour, 3 days after transfection,respectively. MSCs was induced to differentiate into neuron-like cells by-ME. The neural cell specific marker Nestin,NSE,GFAP were determined by immunocytochemistry stain technique. Results Expresion of caveolin-1 vanished by tranfection of caveolin-1 siRNA,survival ratio of MSCs by transfection decreased and there was significant differences between no-transfection and transfection-controlling group after 24 hours and 3 days of transfection(P<0.01).The differentiation efficiency of MSCs transfected by caveolin-1 siRNA into neuron-like cells and the expression of NSE were increased significantly than no-transfection and transfection-controlling group, The percentage of positive NSE was(75.5±2.5)% after 24 hours and(81.6±3.5)% after 6 days, there was no expression of GFAP. There was significant differences between no-transfection and transfection-controlling group(P<0.01). Conclusions The differentiation efficiency of MSCs increased by suppressing caveolin-1 expression.
