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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。
OBJECTIVE: To observe the effect of CXCL12 on the secretion of γ-aminobutyric acid (GABA) and acetylcholine in neurons of hippocampus cultured in vitro and to explore its receptor mechanism. Methods: The intracellular localization of CXCL12 and its receptor CXCR4 were observed by immunofluorescence staining. The secretion of CXCL12 in hippocampal neurons and the secretion of neurotransmitter in hippocampal neurons were detected by ELISA , And changes in neurotransmitter release following CXCR4 blockade with CXCR4 blocker (AMD3100). Results: CXCL12 mainly distributed in the cytoplasm of hippocampal neurons, and was also expressed on the cell membrane. CXCR4 was mainly expressed on the cell membrane of hippocampal neurons. During the in vitro culture of hippocampal neurons, 5, 7 and 10 d), the secretion of CXCL12 decreased gradually. After treated with 50 ng / ml CXCL12 for 48 hours, the secretion of GABA and acetylcholine appeared different changes with the prolongation of culture time, and the secretion of acetylcholine While the secretion of GABA showed a transient decline and a slight transient increase; by blocking its receptor CXCR4 by AMD3100, the secretion of acetylcholine decreased, but the secretion of GABA increased. Conclusion: CXCL12 induces the increase of acetylcholine secretion and the decrease of GABA secretion through CXCR4 receptor. These phenomena indicate that CXCL12 can stimulate the secretion of GABA and acetylcholine as an important nerve growth factor.