CAJ | 학술논문

目的探索灯盏花素注射液体外诱导大鼠骨髓间充质细胞(BMSCs)分化为神经元和胶质细胞的可行性.方法贴壁法分离纯化SD大鼠骨髓间充质细胞.第4代细胞行表型鉴定后,用灯盏花素注射液诱导,每6h倒置相差显微镜观察形态变化,免疫细胞化学染色鉴定诱导后细胞的神经元特异性稀醇化酶(NSE)、神经胶质纤维酸性蛋白(GFAP)的表达情况,四甲基偶氮唑盐(MTT)检测不同浓度灯盏花素注射液诱导后细胞的活力,流式细胞术及RT-PCR检测诱导前后细胞中NSE、GFAP mRNA的表达变化.结果 BMSCs表型鉴定为CD44~+、CD54~+、CD34~-,诱导18h后BMSCs胞体开始收缩,有突起伸出,24h后突起增多形成网络结构.免疫细胞化学染色,NSE阳性表达率为(48.7±3.4)%,GFAP阳性表达为(56.8±4.2)%,流式细胞仪检测诱导24h后的细胞NSE及GFAP蛋白表达量均较未诱导组升高,RT-PCR检测诱导后细胞表达NSE、GFAP mRNA,未诱导的细胞则不表达.结论灯盏花素注射液可诱导大鼠骨髓间充质细胞在体外分化为神经元和神经胶质细胞.
목적 탐색등잔화소주사액체외유도대서골수간충질세포(BMSCs)분화위신경원화효질세포적가행성.방법 첩벽법분리순화SD대서골수간충질세포.제4대세포행표형감정후,용등잔화소주사액유도,매6h도치상차현미경관찰형태변화,면역세포화학염색감정유도후세포적신경원특이성희순화매(NSE)、신경효질섬유산성단백(GFAP)적표체정황,사갑기우담서염(MTT)검측불동농도등잔화소주사액유도후세포적활력,류식세포술급RT-PCR검측유도전후세포중NSE、GFAP mRNA적표체변화.결과 BMSCs표형감정위CD44~+、CD54~+、CD34~-,유도18h후BMSCs포체개시수축,유돌기신출,24h후돌기증다형성망락결구.면역세포화학염색,NSE양성표체솔위(48.7±3.4)%,GFAP양성표체위(56.8±4.2)%,류식세포의검측유도24h후적세포NSE급GFAP단백표체량균교미유도조승고,RT-PCR검측유도후세포표체NSE、GFAP mRNA,미유도적세포칙불표체.결론 등잔화소주사액가유도대서골수간충질세포재체외분화위신경원화신경효질세포.
Objective To explore the feasibility of differentiation into neural cells from bone marrow stromal cells(BMSCs) in vitro by breviscapine,and to offer reference for the applying of BMSCs. Methods The bone marrow stromal cells of SD rat were separated and purified by passsage culture.After phenotype characterization,the 4th passage of BMSCs was exposed to breviscapine. Differentiation was observed under phase contrast microscope every 6 hours and the cells were stained immunocytochemically with neuronspecific enolase(NSE)and glial fibrillary acidic protein(GFAP).The induced cells activity was detected by MTT. NSE and GFAP were determined by flow cytometry and RT-PCR. Results The BMSCs were CD44~+,CD54~+,CD34~-. After 18 hours of induction, cytoplasm of BMSCs partly contracted with protruding;The immunocytochemical staining was performed on cells after induction for 24 hours,the rates of NSE and GFAP staining positive were(48.7±3.4)% and(56.8±4.2)%.By using flow cytometer, expressions of NSE and GFAP showed much higher in induced group than that in control group. By using RT-PCR, expressions of NSE and GFAP were positive in induced group. Conclusion Breviscapine could induce adult rat BMSCs to differentiate into neuron and glial cells.