糖皮质激素致星形胶质细胞内钙浓度快速升高.doc

糖皮质激素致星形胶质细胞内钙浓度快速升高 　 作者：李茂全，王艳艳，杨书，陈朝琼，杨晓虹，余争平 【关键词】 应激；糖皮质激素；星形细胞；钙 　　Rapid elevation of intracellular free calcium concentration in astrocytes after treated by glucocorticoid 【Abstract】 AIM: To explore the mechanisms of glucocorticoid (GC) leading to the elevation of intracellular free calcium concentration (［Ca2+］i) in astrocytes in early effect phase. METHODS: A cell culture model of neonatal Wistar rat hippocampal astrocytes was used. After exposed to corticosterone (CORT) (1 μmol/L), NmethylDaspartate (NMDA), MK801 and BSACORT, respectively, changes of ［Ca2+］i in astrocytes were observed by laser confocal microscope. RESULTS: CORT induced rapid elevation of intracellular Ca2+ fluorescence intensity within 10 min （177.1±3.8， Plt;0.01) compared with controls (141.2±4.7), but induced no significant change when there was no Ca2+ out of the cell （148.5±6.3). NMDA caused further increase of intracellular Ca2+ after pretreatment with CORT (195.3±7.2， Plt;0.01). MK801 partially suppressed the prolonged intracellular Ca2+ elevation (186.4±1.8). BSACORT had almost the same effect as the CORT treatment （200.7±12.2, Plt;0.01). CONCLUSION: The rapid effect of GC induces significant ［Ca2+］i elevation in astrocytes during stress, and maybe NMDA receptor involves in this process. 【Keywords】 stress; glucocorticoid; astrocytes; calcium 【摘要】 目的：研究应激水平的糖皮质激素（GC）在早期快速效应中对星形胶质细胞内游离Ca2+浓度（［Ca2+］i）的影响及作用机制. 方法：原代培养新生Wistar大鼠海马星形胶质细胞，用1 μmol/L的皮质酮(CORT)处理星形胶质细胞，激光共聚焦显微镜下观察CORT快速作用致星形胶质细胞内游离［Ca2+］i的变化，并设立N甲基D天冬氨酸(NMDA)，受体拮抗剂5甲基二氢丙环庚烯亚胺马来酸(MK801)，牛血清白蛋白偶联CORT （BSACORT）及无细胞外钙等干预组,进一步检测［Ca2+］i变化. 结果：与未处理对照细胞（141.2±4.7）相比，CORT可快速介导星形胶质细胞内荧光强度增加（177.1±3.8， Plt;0.01)，但细胞外钙为零时无明显变化（148.5±6.3）；NMDA受体激活可使荧光强度进一步增强(195.3±7.2，Plt;0.01)，NMDA受体拮抗剂MK801可部分拮抗荧光强度升高(186.4±1.8)；BSACORT可产生与CORT类似的效应（200.7±12.2，Plt;0.01). 结论：应激水平的GC在早期可快速增加星形胶质细胞胞内［Ca2+］i，激活NMDA受体是其中的重要机制之一. 【关键词】 应激；糖皮质激素；星形细胞；钙 星形胶质细胞作为中枢神经系统（central nervous system, CNS）内数量最多的细胞，广泛参与CNS内信息的处理与传导，并在神经退行性疾病、脑老化及外伤、炎症等病变的发生发展中扮演重要角