【摘要】 目的:观察CXCL12对体外培养的海马神经细胞神经递质γ-氨基丁酸(GABA)和乙酰胆碱分泌的影响,并探讨其受体机制。方法:通过免疫荧光组织化学染色方法观察CXCL12和其受体CXCR4在细胞内的定位;ELISA方法分别检测海马神经细胞在体外培养过程中CXCL12的分泌变化、CXCL12对海马神经细胞神经递质分泌的影响、以及用CXCR4阻断剂(AMD3100)阻断CXCR4后神经递质释放的变化。结果:CXCL12主要分布在海马神经细胞的细胞浆内,细胞膜上也有表达,其受体CXCR4主要表达在海马神经细胞的细胞膜上;在海马神经细胞体外培养过程中,随时间延长(1、3、5、7、10 d),CXCL12的分泌逐渐减少;采用50 ng/ml的CXCL12作用于海马神经细胞后,随着培养时间的延长,GABA和乙酰胆碱的分泌出现不同的变化,其中乙酰胆碱的分泌逐渐增加,而GABA的分泌则呈现一过性下降又短暂轻微升高的趋势;通过AMD3100阻断其受体CXCR4,则乙酰胆碱分泌减少,但GABA的分泌增加。结论:CXCL12通过CXCR4受体引起乙酰胆碱分泌增加,GABA分泌减少。这些现象表明CXCL12可以作为一种重要的神经生长因子刺激GABA和乙酰胆碱的分泌。更多还原

【Abstract】 Objective: To investigate the effects of CXCL12 on the secretion of GABA and ACh in cultured hippocampal cells in vitro and explore its receptor mechanisms as well.Methods: Locations and expressions of CXCL12 and its receptor CXCR4 were determined by immunofluorescence histochemistry.In addition,the secretion change of CXCL12 on the process of culture in vitro and the effects of CXCL12 on the secretion of GABA and ACh,the change of culture cells secreting GABA and ACh after using CXCR4 blocking agent-AMD3100 were detected by ELISA method,respectively.Results: CXCL12 is mainly located in cytoplasm,a little is observed in cell membrane.On the other hand,CXCR4 is mainly located in cell membrane.In addition,CXCL12 secretion increased gradually with culture time extension(1,3,5,7 and 10 days).When 50 ng/ml of CXCL12 was used to stimulate the cultured hippocampal cells,ACh secretion grew up sharply with culture time extension.However,GABA secretion demonstrated a transient decline and then a short rise.AMD3100 as a kind of CXCR4 blockers halted these progresses by reducing ACh secretion and increasing GABA secretion.Conclusion: CXCL12 may alter GABA and ACh secretion by CXCR4 pathway in cultured hippocampal cells,which may suggest CXCL12 could be considered as an important neurotrophic factor to stimulate GABA and Ach releasing.更多还原