Wnt3a在大鼠骨髓源性胆碱能神经元中的表达  

Expression of Wnt3a in bone marrow stromal cells derived cholinergic neurons

作　　者：王晋丽[1] 王春芳[1] 王晓霞[1] 李鹏飞[1] 李静[1] 蔚洪恩[1] 索耀君[1] 王菲[1] 

机构地区：[1]山西医科大学医学实验中心分子生物学实验室,太原030001

出　　处：《山西医科大学学报》2008年第1期7-10,95,共5页Journal of Shanxi Medical University

摘　　要：目的通过研究大鼠骨髓基质细胞诱导的胆碱能神经元中Wnt3a的表达变化,了解Wnt3a在胆碱能神经元诱导分化过程中的作用,揭示神经元分化过程中可能的信号通路。方法取Wistar雄性大鼠的股骨骨髓,培养骨髓基质细胞,经三代纯化CD71免疫细胞化学鉴定后进行分组实验。实验共分四组:骨髓基质细胞组,未加任何诱导因子;胆碱能神经元诱导分化组,加入Shh,RA和bFGF诱导基质细胞向胆碱能神经元分化,并用ChAT鉴定胆碱能神经元;神经干细胞诱导组,加入bFGF、EGF和RA诱导基质细胞为神经干细胞;自然分化组,加入血清诱导分化。提取细胞总RNA,利用RT-PCR进行Wnt3a的半定量分析。结果与对照的骨髓基质细胞组,神经干细胞诱导组及自然分化组相比,Wnt3a在胆碱能神经元组中表达最高并且有统计学差异。结论Wnt3a在骨髓基质细胞来源的胆碱能神经元中高表达,提示Wnt3a可能是胆碱能神经元分化的Wnt信号通路中的一个环节。Objective To study the expression of Wnt3a in cholinergic neurons differentiated from bone marrow stromal cells （BM- SCs） and to explore the possible signal pathway during neuron differentiation. Methods Bone marrow was dissected from Wistar male rat. BMSCs were cultured and identified after purification for three passages by CD71 immunocytochemistry. Then BMSCs were divided into four groups： BMSCs group, without any induction factors; cholinergic neurons induced group; neural stem cells （NSCs） induced group; natural differentiation group, induced with bovine serum. In cbolinergic neurons induced group, Shh, RA and bFGF were used to induce BMSCs to differentiate into cholinergic neurons and examined by ChAT antigen, while in NSCs induced group, bFGF, EGF and RA were used to induce BMSCs to differentiate into NSCs. Total RNA was extracted and Wnt3a was analyzed by reverse-transeription PCR. Results Compared with BMSCs group, NSCs induced group and natural differentiation group, Wnt3a expression was higher in cholinergic neurons（ P 〈 0.01 ）. Conclusion Wnt3a has a high expression in BMSCs original cholinergic neurons, which indicate that Wnt3a may be a possible node in wnt signal passageway during cholinergic neurons differentiation.

关 键 词：骨髓基质细胞 诱导 分化 胆碱能神经元 WNT3A 

分 类 号：Q786[生物学—分子生物学]