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第三军医大学大坪医院野战外科研究所
纸质出版日期:2008,
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[1]张云东,顾小红,朱佩芳,王正国,周继红,许民辉.促肾上腺皮质激素释放激素对下丘脑神经元CREB的调控[J].重庆医科大学学报,2008(09):1034-1037.
ZHANG Yun-dong, et al. Regulation of P-CREB signals in rat hypothalamic neurons by exogeneous corticotropin—releasing hormone(CRH)[J]. Journal of Chongqing Medical University, 2008,(9):1034-1037.
目的:研究促肾上腺皮质激素释放激素(Corticotropin-releasing hormone
CRH)对下丘脑神经元内CREB(CAMP-responsive element-binding protein)含量变化的调节作用。方法:取孕17d胎鼠下丘脑分散培养下丘脑神经元
采用PTI(Photon technology international
PTI)荧光成像系统测量下丘脑神经元胞浆内游离钙浓度([Ca2+]i)变化;测量和分析外源性CRH对培养下丘脑神经元内钙信号变化的影响
用Western blot方法测定神经元内P-CREB的含量变化
分别与用细胞膜L-型钙通道拮抗剂尼莫地平或CRH1受体(CRH1R)特异性拮抗剂CP-154526处理下丘脑神经元后[Ca2+]i和P-CREB的变化作比较。结果:正常对照组的下丘脑神经元[Ca2+]i含量较低
外源性CRH刺激后
[Ca2+]i立即升高;预先应用尼莫地平或CP-154526处理再用CRH刺激的神经元[Ca2+]i含量的增加明显被抑制
同时也可明显抑制神经元内P-CREB含量的增加。结论:在急性应激反应中
CRH可直接与下丘脑神经元CRH1R结合
开放膜L-型钙离子通道促使钙离子内流使[Ca2+]i明显增加
从而进一步激活神经元内P-CREB信号通路。说明在调节下丘脑神经元激活过程中CRH可能起了重要的作用。
Objective:To study the function and regulation of CREB signaling in rat hypothalamic neurons by exogeneous corticotropin-releasing hormone(CRH). Methods:Perifused primary cultures of rat hypothalamic neurons. Intracelluar free Ca2+ concentr ation([Ca2+]i) was assayed at 32℃ by fluorescence analysis
the coverslip was mounted in a Delta Scan spectrofluorimeter(Photon Technology International);and phospho-CREB by western blot. Results:The basal lower intracellular Ca2+ level of perifused primary cultures of rat hypothalamic neurons cultured on coverslipy was markedly increased by CRH
and significantly attenuated by application of nifedipine(Nif) or CP-154526. CRH may cause the increase of phospho-CREB in the neuron of hypothalamus
while CP-154526
Nif can significantly inhibit the production of phospho-CREB. Conclusion: CRH may excite hypothalamic neurons directly suggest that phospho-and[Ca2+]i
suggesting a role of CRH type 1 receptor in the activation of CRH gene expression.
钙通道下丘脑神经元P-CREB促肾上腺皮质激素释放激素
Calcium signalingPhosphorylated cAMP-responsive element-binding proteinHypothalamicCorticotropin-releasing hormone
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