摘要
目的:探讨海马胆碱能神经元刺激肽(hippocampal cholinergic neurostimulating peptide,HCNP)与海马提取液对海马神经干细胞向神经元及胆碱能神经元分化的协同作用。方法:取孕17 d SD大鼠海马组织进行神经干细胞的培养、增殖、传代,并将其接种于2块24孔板中,培养基中均含B27、微量脑源性神经营养因子(BD-NF),将培养孔分为6组:H+T组,加入HCNP和切割穹窿海马伞侧海马提取液;H+N组,加入HCNP和正常侧海马提取液;H组,加入HCNP;T组,加入切割穹窿海马伞侧海马提取液;N组,加入正常侧海马提取液;对照组,不加HCNP和海马提取液。分化14 d后行MAP-2/ChAT免疫荧光双标检测。结果:与其它各组相比,H+T组分化的MAP-2阳性神经元最多,ChAT/MAP-2双标阳性神经元占MAP-2阳性神经元的比例最高(P<0.05);与N组相比,H+N组MAP-2阳性神经元较多,ChAT/MAP-2双标阳性神经元占MAP-2阳性神经元的比例也较高(P<0.05);与对照组相比,H组T组均可见一定数量的MAP-2阳性神经元及ChAT/MAP-2双标阳性神经元(P<0.05);N组以及对照组中MAP-2阳性神经元的数量和ChAT/MAP-2双标阳性神经元占MAP-2阳性神经元的比例则最低。结论:在微量BDNF存在的情况下,HCNP可促进海马神经干细胞向神经元和胆碱能神经元分化,切割侧海马提取液与HCNP可发挥协同作用。
Objective： To investigate the effects of hippocampal cholinergic neurostimulating peptide（HCNP） on the neuronal or cholinergic differentiation of hippocampal neural stem cells（NSCs） in vitro.Methods： The E17 hippocampus were dissociated and plated in culture flasks with NSC specific medium.After proliferation and passage,the NSCs were plated into two 24-well cultured plates with medium containing B27 and brainderived neurotrophic factor（BDNF）.The wells were divided into 6 groups： group H＋T with HCNP and transected hippocampal extracts;group H ＋ N with HCNP and normal hippocampal extracts;group H with HCNP;group T with transected hippocampal extracts;group N with normal hippocampal extracts;group control without HCNP and any extracts.At 14 day after incubation,the cells were subjected to immunofluorescence against MAP-2 and ChAT antibodies.Results： Immunofluorescence showed that compared with other groups,there were much more MAP-2 positive neurons in the group H＋T,most of which were double positive to ChAT and MAP-2（P0.05）.Compared with the group N,more MAP-2 positive neurons and ChAT/MAP-2 double positive neurons were found in group H＋N（P0.05）.There were also more MAP-2 positive or ChAT/MAP-2 double positive neurons in the groups T and H than the group control（P0.05）.Among all of the groups,the numbers of MAP-2 or ChAT/MAP-2 positve neurons in N and control groups were least.Conclusion： HCNP can promote hippocampal NSCs to differentiate into neurons and cholinergic neurons in the presence of BDNF with low concentration,and HCNP treatment combined with the fimbria/fornix-transection hippocampal extracts has synergistic effects on neuronal differentiation of hippocampal NSCs.
出处
《神经解剖学杂志》
CAS
CSCD
北大核心
2011年第5期539-544,共6页
Chinese Journal of Neuroanatomy
基金
江苏省高校优势学科建设工程资助项目(PAPD)
南通市科技应用研究计划(K2009025)
江苏省高校自然科学研究项目(09KJB310011)
