Lhx8基因沉默抑制NGF诱导的海马NSCs向胆碱能神经元分化的研究  

Lhx8 gene silence inhibits NGF induced cholinergic differentiation of hippocampal neural stem cells( NSCs) in vitro

作　　者：李浩明[1] 金国华[1] 朱培培[1] 施金洪[1] 邹琳清[1] 张新化[1] 田美玲[1] 衣昕[1] 秦建兵[1] 成翔[1] 

机构地区：[1]南通大学医学院人体解剖学系,江苏省神经再生重点实验室,南通226001

出　　处：《神经解剖学杂志》2014年第6期629-633,共5页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(31070937);江苏省高校自然科学研究项目(13KJB310011);江苏高校优势学科建设工程资助项目(PAPD)

摘　　要：目的:明确Lhx8在神经生长因子(nerve growth factors,NGF)促进海马神经干细胞向胆碱能神经元分化中的作用。方法:体外培养海马神经干细胞并应用NGF促进其向胆碱能神经元分化的过程中,将构建的Lhx8干扰慢病毒加入至培养液中,7 d后应用免疫荧光标记技术检测分化所得胆碱乙酰转移酶(choline acetyltransferase,Ch AT)和微管相关蛋白-2(microtubule-associated protein 2,MAP-2)双标阳性细胞。结果:在空白对照组中,仅检测到少量的Ch AT/MAP-2双标阳性细胞;在NGF组或是NGF联合阴性对照慢病毒组中则可检测到较多的Ch AT/MAP-2双标阳性细胞;在加入Lhx8干扰慢病毒的实验组中,分化所得的Ch AT/MAP-2双标阳性细胞数较NGF组或是NGF联合阴性慢病毒组明显减少。结论:Lhx8基因沉默抑制了NGF诱导的海马神经干细胞向胆碱能神经元的分化。Objective：To understand possible effects of Lhx8 in nerve growth factors（ NGF）-induced cholinergic neuronal differentiation of hippocampal neural stem cells（ NSCs） in vitro.Methods：Hippocampal NSCs were cultured in vitro and NGF was used to promote cholinergic neuronal differentiation,the Lhx8 interference lentiviral vectors（ Lv-Lhx8-RNAi） were added into culture medium.7 d later,choline acetyltransferase（ Ch AT） and microtubule-associated protein2（ MAP-2） double-labeled cells were detected by using immunofluorescence.Results：In control group,only a little Ch AT / MAP-2 double-labeled cells could be detected;in NGF group or NGF combined negative control group,more Ch AT / MAP-2 double-labeled cells were detected;after Lv-Lhx8-RNAi transfection,the number of Ch AT / MAP-2 doublelabeled cells in Lv-Lhx8-RNAi group was less than NGF or NGF combined with negative control group.Conclusion：Lhx8 gene silence inhibited NGF-induced cholinergic neuronal differentiation of hippocampal NSCs in vitro.

关 键 词：Lhx8 基因沉默 神经生长因子 神经干细胞 胆碱能神经元 海马 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]