缺氧对新生大鼠海马神经干细胞增殖和分化的影响  

Influence of Hypoxia on Proliferation and Differentiation of Neural Stem Cells from Hippocampus of Neonate Rats

作　　者：赵久红[1] 崔燕[2] 唐源远[1] 季丽莉[1] 王立芹[3] 王振宇[1] 

机构地区：[1]中国医科大学基础医学院人体解剖学教研室,辽宁沈阳110001 [2]中国医科大学,7年制91期4班辽宁沈阳110001 [3]聊城职业技术学院基础部人体解剖学教研室,山东聊城252000

出　　处：《解剖科学进展》2009年第2期168-171,共4页Progress of Anatomical Sciences

基　　金：辽宁省自然科学基金项目(N0.20082097)

摘　　要：目的探讨单纯性缺氧对新生大鼠海马神经干细胞(NSCs)增殖和分化的影响。方法将36只新生1d大鼠随机均分为N组(正常对照组)、Q2组(缺氧2h组)、Q5组(缺氧5h组)。将海马组织在无血清培养液(均含有碱性成纤维生长因子bFGF、表皮生长因子EGF和白细胞抗原B27)中,分别进行原代培养、单克隆培养和传代培养,单克隆培养细胞进行巢蛋白(Nestin)和Hoechst33342免疫荧光双标染色;传代培养的细胞传三代诱导分化,诱导分化3d的细胞分别行神经元特异性烯醇化酶(NSE)、胶质纤维酸性蛋白(GFAP)和Hoechst3342免疫荧光双标染色,计数NSE阳性细胞比例,进行统计学分析。结果单克隆培养的细胞均呈Nestin阳性表达,诱导分化的细胞部分呈NSE或GFAP阳性表达。Q2组、N组、Q5组细胞原代培养克隆球直径达2μm的时间分别为7d、10d、13d。传3代后,对各组细胞进行诱导分化,Q2组细胞诱导分化为神经元的比例明显高于N组,Q5组的细胞诱导分化为神经元的比例明显低于N组。结论短时间缺氧可促进在体海马神经干细胞增殖和诱导其向神经元分化,长时间缺氧则抑制在体神经干细胞增殖和向神经元分化。Objective To study the influence of hypoxia on the proliferation and differentiation of neural stem cells（NSCs） in hippocampus of neonate rats. Methods Neonate rats were divided into： N group （normal rats）, Q2 group （exposed to 92%N2 and 8%02 for 2h）, Q5 group （exposed to 92%N2 and 8%02 for 5h）. Then NSCs in hippocampus of neonate rats were cultured in the serum-free medium containing basic fibroblast growth factor （bFGF）, epidermal growth factor（EGF）and B27. Cells were monocloned and induced to be differentiated by fetal bovine serum for 3 days. Immunofluorescence was used to identify the nestin positive cells of sigle clone NSCs and neuron specific enolase （NSE） positive and glial fibrillary acidic protein （GFAP） positive in 3G NSCs differentiated for 3 days. Results The monoclonal cells were all nestin positive,part of induced and differentiated cells were either NSE positive or GFAP positive. The percentage of NSE positive cells differentiated from NSCs was much higher in Q2 group than in N group and Q5 group （P〈0.05） and lower in Q5 group than in N group. Conclusion Short-time hypoxia increased the percentage of NSE positive cells differentiated from NSCs in the hippocampus of neonate rats transiently.

关 键 词：缺氧 新生大鼠 海马 神经干细胞 分化 

分 类 号：R338.26[医药卫生—人体生理学]