ERK1/2信号转导通路对骨髓基质细胞条件培养液诱导大鼠中脑神经干细胞分化作用的影响  

Influence of ERK1/2 signal-transduction pathway on differentiation of mesencephalic neural stem cells induced by bone marrow stromal cell-conditioned medium

作　　者：顾平[1] 王彦永[1] 马芹颖[1] 温雅[1] 崔冬生[1] 刘力[1] 王铭维[1] 张振清[1] 

机构地区：[1]河北省脑老化与认知神经科学实验室

出　　处：《第二军医大学学报》2007年第6期589-592,共4页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(30400138);河北省自然科学基金(C2004000583).~~~~

摘　　要：目的:探讨ERK1/2信号转导通路在骨髓基质细胞条件培养液(BMSCs-CM)诱导大鼠中脑神经干细胞(NSCs)分化中的作用。方法:将神经干细胞球种植干预先包被多聚赖氨酸的35 mm的培养皿中.神经干细胞球约30 min后贴壁.自然分化组将培养液换为含2%B27的neurobasal培养液,对照组换为BMSCs-CM.抑制剂组将培养液换为含有信号转导通路抑制剂PD98059(5μmol/L)的BMSCs-CM.7 d后采用免疫荧光染色方法观察NSCs后代细胞中神经元和星形胶质细胞的数量比例与形态差别。结果:用成年大鼠BMSCs-CM体外培养新生大鼠中脑NSCs.自然分化组NSCs分化的神经元为(15.7±10.3)%[低干对照组的(51.17±10.30)%.P<0.05],星形胶质细胞为(66.9±14.8)%[明显高于对照组的(23.26±5.60)%.P<0.05]。而抑制剂组神经元所占比例为(39.48±7.29)%[明显低于对照组.P<0.01].星形胶质细胞的比例为(31.01±8.96)%[明显高干对照组,P<0.01]。结论:BMSCs-CM可以提高神经干细胞分化为神经元的比例.并且同时降低了其分化为星形胶质细胞的比例,这两种作用很可能是通过ERK1/2信号转导通路实现的。Objective：To investigate the effect of the ERK1/2 signal-transduction pathway on differentiation of rat mesencephalic neural stem eells（NSCs） in presence of bone marrow stromal cell-conditioned medium（BMSCs-CM）in vitro. Methods： NSCs were cultured on poly-L-lysine （PLL） coated 35 mm dishes. The neurospheres adhered to the bottom 30 min later; then the culture medium of natural differentiation group was replaced by neurobasal medium containing 2% B27; the control group was replaced with BMSCs-CM; and the inhibition group was replaced with BMSCs-CM containing PD98059 （5 μmol/L）. The proportions of neurons and astroeytes and their morphological difference were observed by immunocytochemistry technique 7 d later. Results： We successfully cultured NSCs from adult rat BMSCs-CM. The proportion of neurons in the natural differentiation group （15.70±10.3）% was lower than that in the control group （[51.174±10.30]%, P〈0.05） ;the proportion of astrocytes in the natural differentiation group （66.94±14.8）% was higher than that in the control group （E23.26±5.60]%/,P〈0.05）. The proportion of neurons in the inhibition group （39.48±7.29）% was lower than that in the control group （[51.17±10.30] %, P〈0.05）; the proportion of astrocytes in the inhibition group （31.01±8.96）% was higher than that in the control group （[23.26±5.60]% ,P〈0.01）. Conclusion： BMSCs-CM can up-regulate the proportion of neurons in cultured NSCs and down-regulate the proportion of astrocytes, which might be mediated by the ERK1/2 signal-transduction pathway.

关 键 词：ERKl/2信号转导通路 骨髓基质细胞 条件培养液 神经干细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]