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目的:研究促肾上腺皮质激素释放激素(CRH)对原代培养海马神经元谷氨酸诱发电流(IGLU)的调节作用,并初步探讨其可能机制。方法:应用免疫荧光细胞化学方法观察原代培养的大鼠海马神经元CRH两型受体的表达;通过全细胞膜片钳技术研究CRH对海马神经元IGLU的调节作用及其可能涉及的胞内信号通路。结果:CRH作用2min明显抑制海马神经元IGLU,且呈剂量依赖性;细胞外给予CRH受体广谱拮抗剂α-helical CRH或CRHR1特异性拮抗剂antalarmin能完全阻断CRH对IGLU的抑制作用,CRHR2特异性拮抗剂astressin-2B对此没有明显作用;细胞内给予蛋白激酶C(protein kinaseC,PKC)的抑制剂G6976可阻断CRH对IGLU的抑制作用。结论:多肽类激素CRH可抑制原代培养的海马神经元IGLU,这种作用由CRHR1介导,并可能涉及PKC信号通路。
AIM: To investigate the regulatory effect of corticotropin-releasing hormone (CRH) on glutamate-induced currents (IGLU) in primary cultured hippocampal neurons and its possible mechanism. Methods: Immunofluorescence cytochemistry was used to observe the expression of CRH type 2 receptor in primary cultured hippocampal neurons. Whole cell patch clamp technique was used to investigate the regulatory effect of CRH on hippocampal neurons IGLU and its possible intracellular signal path. Results: CRH inhibited the IGLU of hippocampal neurons in a dose-dependent manner at 2min. The α-helical CRH or CRHR1-specific antagonist antalarmin could block the inhibitory effect of CRH on IGLU completely. CRHR2-specific antagonist astressin-2B has no significant effect on this; intracellular administration of protein kinase C (PKC) inhibitor G6976 can block the inhibitory effect of CRH on IGLU. CONCLUSION: Polypeptide hormone CRH can inhibit the primary culture of hippocampal neurons IGLU, which is mediated by CRHR1 and may involve PKC signaling pathway.