皮质酮对原代培养的海马神经元及其[Ca^(2+)]_i和钙/钙调蛋白依赖性蛋白激酶Ⅱ表达的影响  

Effects of corticosterone on primarily cultured hippocampal neurons,their [Ca^(2+)]_i and CaMKⅡ expression

作　　者：孙臣友[1] 戚双双[2] 陈朝玉[1] 刘能保[3] 张敏海[3] 

机构地区：[1]温州医学院解剖学教研室,温州325035 [2]温州医学院附属第二医院药剂科,温州325035 [3]华中科技大学同济医学院组织学与胚胎学教研室,武汉430030

出　　处：《解剖学杂志》2006年第3期270-274,共5页Chinese Journal of Anatomy

摘　　要：目的:探讨皮质酮(CORT)对培养的海马神经元及其[Ca2+]i和钙/钙调蛋白依赖性蛋白激酶Ⅱ(CaMKⅡ)表达的影响和可能的机制。方法:海马神经元被分为不同终浓度的CORT处理组、CORT+MK-8 0 1或高浓度葡萄糖组。采用MTT法、流式细胞术、荧光标记和免疫细胞化学法,观察海马神经元活力、死亡方式[、Ca2+]i和CaMKⅡ表达的变化规律。结果:1 0-6和1 0-5mol/L CORT组,其海马神经元的活力明显降低,分别以凋亡和坏死为主,并使海马神经元[Ca2+]i显著升高;CaMKⅡ的表达明显减少。MK-8 0 1和高浓度葡萄糖均能拮抗1 0-6mol/L CORT对海马神经元的损伤作用。结论:在CORT的作用下,海马神经元发生凋亡和坏死;[Ca2+]i升高可能既是海马神经元损伤的结果,又是引起其发生凋亡和CaMKⅡ表达下降的原因。Objective： To explore the effects of corticosterone （CORT） on cultured hippocampal neurons and their [Ca^2＋]i and calcium/calmodulin-dependent protein kinase Ⅱ （CaMK Ⅱ ） expression. Methods： Hippocampal neurons were divided into groups according to different concentrations of CORT, CORT with MK-801 or high concentration of glucose. MTT, flow cytometry, fluorescent labeling and immunocytochmistry were used to observe the changes of hippocampal neurons cell activity, cell death, and [Ca^2＋] and CaMK Ⅱ expression. Results： Treament with CORT （10^-6mol/L and 10^-Stool/L） decreased cells viability markedly through apoptosis and necrosis. Both MK-801 and high concentration of glucose antagonized the damaging effect of CORT at 10^-6 mol/L on hippocampal neurons. Furthermore, CORT increased [Ca^2＋]i and decreased CaMK Ⅱ expressions significantly in hippocampal neurons. Conclusion： Apoptosis and necrosis occur in the hippocampal neurons treated with CORT. The increase of [Ca^2＋] might be caused by impairment of the hippocampal neurons and lead to the apoptosis of hippocampal neurons and low expressions of CaMK Ⅱ.

关 键 词：皮质酮 海马 神经元 [CA^2+]I 钙/钙调蛋白依赖性蛋白激酶Ⅱ 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]