脑源性神经营养因子诱导大鼠骨髓基质细胞成为神经干细胞及其分化作用的实验研究

Effects of brain derived neurotrophic factor on inducing adult rat bone marrow stromal cells to neural stem cells and their differentiation

目的探讨脑源性神经营养因子(BDNF)诱导大鼠骨髓基质细胞(BMSCs)成为神经干细胞及其分化作用。方法取成年大鼠BMSCs,分别以BDNF和BDNF+RA(维甲酸)作为诱导物诱导,于诱导3d、7d后行巢蛋白(Nestin)、神经元特异烯醇化酶(NSE)、胶质纤维酸性蛋白免疫细胞化学染色。结果诱导3天后BDNF和BDNF+RA诱导组均有大量Nestin染色阳性细胞,BDNF+RA组阳性率高于BDNF组(P<0.01)。NSE、GFAP免疫阳性细胞在诱导3d后也有少量表达。诱导7天后BDNF和BDNF+RA诱导组Nestin阳性细胞明显减少,与诱导3天后比较差异有显著性(P<0.01),而NSE、GFAP阳性细胞数增多,与诱导3天后相比差异有显著性(P<0.01),且BDNF+RA组阳性率高于BDNF组(P<0.01)。结论联合应用BDNF与RA可提高BMSCs神经转化,并促进其向神经元及星形胶质细胞细胞分化。

Objective： To investigate the effects of brain derived neurotrophic factor （BDNF） inducing adult rat bone marrow stromal cells （BMSCs）to the neural stem cells and their descendants. Methods： Purified BMSCs were induced by BDNF alone or by combination of BDNF and refinoic aeid（RA）. In 3 days and 7 days after inducing, nestin , Neuron-specific enolase（NSE） and glial fibrillary acidic protein（GFAP） were detected by immunocytoehemistry. Results： After 3 days of induction by BDNFor combination of BDNF and RA, there are a lot of cells positive for nestin . The percent of cells positive for nestin in combination of BDNF and RA group were significantly increased than BDNF group. NSE and GFAP-positive cells were little increased. After 7 days of induction. Cells positive for nestin in BDNF group or combination of BDNF and RA group, were decreased markedly compared with that 3 days of induction , but NSE and GFAP-positive ceils were increased significantly compared with that 3 days of induction . Furthermore the combination of BDNF and RA group showed a maximal increase . Conclusion.. Combination of BDNF and RA might improve the neural induction of BMSCs ,and then differentiate into neurons and glial cells.