葛根素对骨髓间充质细胞的诱导分化  

The induction and differentiation of puerarin to MSCs in vitro

作　　者：邓仪昊[1] 陈道邦[1] 徐立[1] 史凯[1] 

机构地区：[1]四川大学基础医学与法医学院人体解剖学教研室,成都610041

出　　处：《中国临床解剖学杂志》2008年第1期83-86,共4页Chinese Journal of Clinical Anatomy

基　　金：四川省科学技术厅应用基础研究项目(2006J13-008-4)

摘　　要：目的:探索葛根素体外诱导大鼠骨髓间充质细胞(Bone Marrow Stromal Cells MSCs)分化为神经元和胶质细胞的可行性,为中药在细胞定向分化中的应用提供理论和实验依据。方法:用贴壁法分离纯化SD大鼠骨髓间充质细胞。培养传至第5代,诱导组以2.5mg/ml的葛根素预诱导24h后,用17.5mg/ml的葛根素无血清培养基诱导,未诱导组加入等量培养基,24h后相差显微镜观察形态变化,免疫细胞化学染色鉴定诱导后的细胞神经元特异性稀醇化酶(neuronspecific enolase NSE),神经胶质纤维酸性蛋白(glial fibrillary acidic protein GFAP)蛋白的表达情况,MTT检测不同浓度葛根素诱导后细胞的活力,流式细胞仪及RT-PCR检测诱导前后细胞中NSE、GFAP的表达情况。结果:诱导18h后BMSCs胞体收缩,有突起伸出,24h后突起增多呈网状。免疫细胞化学染色,NSE阳性表达率为(53.3±4.3)%,GFAP阳性表达为(64.5±5.2)%,流式细胞仪检测诱导24h后的细胞NSE及GFAP表达量均较未诱导组升高,RT-PCR检测诱导后细胞表达NSE、GFAP,未诱导的细胞则不表达。结论:葛根素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。Objective： To explore the feasibility of MSCs which induced by puerarin differentiating into neural cells and glial cells, and provide the theoretical and experimental basis for the implement of Traditional Chinese Medicine on the cell orientational differention. Methods： The bone marrow stromal cells of SD rat were separated and purified by passsage culture. The 5th passage of BMSCs was induced by 2.5 mg/ml puerarin for 24 h beforehand and was induced by 17.5 mg/ml. Control group was no puerarin. After 24 h, differentiation was observed under phase contrast microscope and the cells were stained immunocytochemically with neuronsp- cific enolase （NSE）and glial fibrillary acidic protein （GFAP）. The induced cells activity was detected by MTT. NSE and GFAP was determined by flow cell motor and RT-PCR. Results： Partly cytoplasm Of BMSCs contracted with protruding afterl 8 hours of induction; and the reticulate protrude increased in 24 hours. The immunocytochemical staining was performed on cells after induction for 24 hours,the rate of NSE and GFAP staining positive was （53.3±4.3）% and （64.5±5.2）% ,flow cytometer has the same result. Conclusion： Puerarin could induce adult rat BMSCs differentiate into neuron and glial cells.

关 键 词：骨髓间充质细胞 葛根素 神经元 神经胶质细胞 

分 类 号：R285.5[医药卫生—中药学]