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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。
Objective To explore the feasibility of breviscapine injection induced differentiation of rat bone marrow mesenchymal cells (BMSCs) into neurons and glial cells in vitro. Methods SD rat bone marrow mesenchymal cells were isolated and purified by adherence method. The fourth generation of cells after phenotypic identification, with breviscapine injection induced morphological changes observed by inverted phase contrast microscope every 6h, immunocytochemical staining of cells after induction of neuron-specific excitatory enzyme (NSE), glial Fibronectin (GFAP) was detected by MTT assay. The viability of cells induced by breviscapine injection was detected by flow cytometry and RT-PCR. The levels of NSE, GFAP Changes in mRNA expression. Results The phenotypes of BMSCs were identified as CD44 +, CD54 + and CD34-. BMSCs began to shrink after being induced for 18h. Immunocytochemical staining showed that NSE positive expression rate was (48.7 ± 3.4)% and GFAP positive expression was (56.8 ± 4.2)%, respectively. Compared with non-induced group, High, the expression of NSE and GFAP mRNA was detected by RT-PCR, while the non-induced cells were not expressed. Conclusion Breviscapine injection can induce rat bone marrow mesenchymal cells to differentiate into neurons and glial cells in vitro.