摘要
目的 探讨降钙素促进中脑导水管周围灰质（PAG）神经元谷氨酸释放的机制.方法 原代培养PAG神经细胞,免疫组化法鉴定原代培养的PAG神经细胞为PAG神经元细胞;高效液相色谱法（HPLC）观察鲑鱼降钙素（sCT）是否诱发PAG神经元谷氨酸释放并探讨其可能的细胞信号机制.结果 sCT可促进PAG神经元谷氨酸释放（P＜0.01）,此作用可被sCT受体拮抗剂sCT8-32所阻断（P＜0.01）.sCT诱发的PAG神经元谷氨酸释放可以被蛋白激酶C（PKC）阻断剂白屈菜季氨碱（Che）部分阻断（P＜0.01）.结论 sCT通过与PAG神经元上的降钙素受体结合,激活蛋白激酶PKC信号途径引起PAG神经元谷氨酸释放增加.
Objective To investigate the effect and mechanism of calcitonin（CT） in glutamate release from primary cultured neurons of midbrain periaqueductal gray matter（PAG）.Methods Primary dissociated culture of PAG neurons was prepared from neonatal Sprague-Dawley （SD） rats.The cultured cells were divided into 4 groups：control group,salmon calcitonin （sCT） group,CT antagonist group （sCT8-32）,and protein kinase C （PKC） inhibitor chelerythrine（Che） group.Each group was further divided into subgroups representing low,middle,and high levels of drugs.Glutamate release from the cultured PAG neurons evoked by sCT and/or other interfering factors was detected by using high-performance liquid chromatography （HPLC） assay.Results （1） Compared with the control group,sCT group yielded a time-dependent and concentration-dependent glutamate release from the cultured PAG neurons,and the most effective concentration of sCT was 20 nmol/L（P＜0.01）.（2） sCT8-32,a selective antagonist of CT receptor,significantly reversed the effect of 20 nmol/L sCT on glutamate release from cultured PAG neurons,and the most effective concentration was 100nmol/L sCT8-32 （P＜0.01）.（3） Incubation of the cultured neurons with Che inhibited the glutamate release from cultured PAG neurons evoked by 20 nmol/L sCT,and 100 μmol/L Che was most effective（P＜0.01）.Conclusion CT receptors participates in the glutamate release from PAG neurons in which intracellular protein kinase C signaling pathway is involved.
出处
《中华内分泌代谢杂志》
CAS
CSCD
北大核心
2014年第6期511-515,共5页
Chinese Journal of Endocrinology and Metabolism
基金
贵州省贵阳市科技计划项目（2011103）
