遗传性癫痫大鼠海马组织Ca^(2+)/Ca_V1.2/CaM/CaMKⅡ信号通路的异常变化  

The abnormal changes of Ca^(2+) /Ca_V1.2/CaM/CaMK Ⅱ signaling pathway in hippocampus of genetic epileptic rat

作　　者：吕昕瞳[1,2] 封瑞[1] 蔡际群[1] 徐晓雪[3] 马丽华[1] 何桂林[1] 胡慧媛[1] 赵金生[1] 赵美眯[1] 郭凤[1] 

机构地区：[1]中国医科大学药学院药物毒理学教研室,辽宁沈阳110001 [2]辽宁省辽阳市第三人民医院,辽宁辽阳111000 [3]中国医科大学附属一院神经内科,辽宁沈阳110001

出　　处：《中国药理学通报》2013年第1期31-36,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81001429;30270535);辽宁省教育厅资助项目(No L2010573)

摘　　要：目的研究遗传性癫痫大鼠(tremor,TRM)海马组织电压门控性L型钙离子通道α1C亚单位(CaV1.2)、钙调蛋白(CaM)、钙调蛋白依赖性蛋白激酶Ⅱ(CaMKⅡ)和细胞内钙离子浓度([Ca2+]i)的变化情况。方法应用West-ern blot法与免疫荧光双标法检测TRM海马CA1、CA3和DG区CaV1.2、CaM和磷酸化CaMKⅡ(p-CaMKⅡ)的蛋白表达及分布;激光共聚焦显微镜检测TRM海马组织中[Ca2+]i。结果与正常Wistar大鼠相比,TRM海马组织中CaV1.2和CaM的蛋白表达明显升高(P<0.01),而p-CaMKⅡ的蛋白表达明显下降(P<0.01);免疫荧光双标法结果显示:CaV1.2、CaM、p-CaMKⅡ在CA1、CA3区的锥体细胞和DG区的颗粒细胞群表达丰富,同时CaV1.2与CaM、p-CaMKⅡ与CaM在海马各区域均存在共定位;激光共聚焦显微镜检测TRM海马细胞[Ca2+]i明显增强(P<0.01)。结论Ca2+/CaV1.2/CaM/CaMKⅡ通路的异常变化可能参与遗传性癫痫大鼠的癫痫发生与发展。Aim To observe the changes of L-type voltage-dependent calcium channel ctl C-class （Car 1.2 ） , calmodulin （ CaM ）, cahnodulin-depend- ent protein kinase Ⅱ （ CaMK Ⅱ） and intracellular cal- cium concentration （ [ Ca2＋ ]i ） in hippocampus of tremor rats （TRMs）. Methods The protein distribu- tions and expressions of Cavl. 2, CaM and phosphoryl- ated CaMK Ⅱ （p-CaMK Ⅱ ） in TRM hippocampus were detected by double-labeling immunofluorescence and Western blot; [ Ca2＋ ]i was measured by confocal laser scanning microscope. Results Compared to control rats, the protein expressions of Carl. 2 and CaM were increased, and p-CaMK Ⅱ was decreased in TRM hippocampus （P 〈 0. 01 ）. Double-labeled ira-munofluorescence consequences suggested that in TRMs, Cavl. 2, CaM and p-CaMK Ⅱ protein appeared throughout CA1, CA3 and DG regions of hippocam- pus. It was noteworthy that co-localized neurons that expressed Carl. 2 with CaM and p-CaMK lI with CaM were also detected. Furthermore, compared to control rats, intracellular calcium concentrations were in- creased （ P 〈 0.01 ）. Conclusion The abnormal changes of Ca2 ＋/Cavl. 2/CaM/CaMK Ⅱ signaling pathway might be involved in TRMs＇ epileptogenesis.

关 键 词：遗传性癫痫大鼠 海马 电压门控性钙通道 钙调蛋 白 钙调蛋白依赖性蛋白激酶Ⅱ 细胞内钙离子浓度 癫痫 

分 类 号：R-332[医药卫生] R322.81