缝隙连接蛋白CX32在颞叶癫痫患者海马中的表达及生胃酮的抑制作用  

Expression of connexin 32 gap-junction proteins in medically intractable temporal lobe epilepsy in humans

作　　者：秦哲[1] 田庆华[2] 姚丽芬[3] 

机构地区：[1]齐齐哈尔医学院,黑龙江齐齐哈尔161006 [2]哈尔滨市解放军第211医院神经内科,黑龙江哈尔滨150086 [3]哈尔滨医科大学附属第一医院神经内科,黑龙江哈尔滨150001

出　　处：《中风与神经疾病杂志》2013年第9期804-808,共5页Journal of Apoplexy and Nervous Diseases

摘　　要：目的观察缝隙连接蛋白(connexin,Cx)32在颞叶癫痫患者手术切除的海马组织中的表达以及缝隙连接阻滞剂生胃酮(carbenoxolone,Carb)和传统抗癫痫药丙戊酸钠(Valproic acid,VPA)对两者表达的影响,探讨缝隙连接在癫痫的发生、发展机制中的作用,为颞叶癫痫的治疗寻找新的方向。方法 (1)以14例颞叶癫痫患者手术切除的海马组织为研究对象,采用免疫组化和Western-blot半定量分析方法检测缝隙连接蛋白(connexin,Cx)Cx32的表达;(2)取颞叶癫痫患者术中切除的海马组织制备成400?m海马脑片,浸入生胃酮液、丙戊酸钠及人工脑脊液中,于9h分别行Western-blot分析,检测中CX32蛋白表达情况。结果免疫组化结果可见:缝隙连接蛋白Cx32在正常对照组海马组织中有比较低水平的表达,颞叶癫痫患者海马组织中Cx32表达明显增强,两者之间有统计学差异(P<0.05)。且CX32主要在神经元的胞体及轴突以及小胶质细胞中表达明显。脑片的Western-blot结果可见:颞叶癫痫患者组及癫痫+丙戊酸钠组的海马组织中Cx32的表达明显增强,两者之间无统计学差异(P>0.05);生胃酮显著抑制了Cx32的表达,对照组与癫痫+生胃酮组两组缝隙连接蛋白Cx32的表达无统计学差异(P>0.05)。而癫痫组、癫痫+丙戊酸钠组两组较癫痫+生胃酮组、对照组缝隙连接蛋白Cx32的表达明显增高,有统计学意义(P<0.05)。结论缝隙连接蛋白Cx32在难治性颞叶癫痫患者海马组织中表达增高,缝隙连接蛋白阻滞剂能够明显降低癫痫病灶中Cx32的表达,且传统抗癫痫药物并不能对缝隙连接蛋白的表达产生影响,缝隙连接的作用机制独立于传统的化学性突触的的作用通路。Objective To compare the level of expression of Cx32 in hippocampi from epileptic and nonepileptic pa- tients, to observe the influence of Carb and VPA on both of them, to assess whether an alteration of gap junction expression in epileptic tissue plays a role in seizure origin and propagation. Methods （ 1 ） This study was performed on hippocampal tissue obtained from 14 patients suffering from pharmacoresistant temporal lobe epilepsy. Cx32 expressions were assessed by both Western-blot and immunohistochemistry methods. （2） Hippocampal specimens from surgery were immediately trans- ferred into cold sucrose solution, Horizontal slices （4004p^m） were cut at 4~C in sucrose solution. Hippocampal specimens were immediately immersed in sucrose solution of VPA and Carla for 9 hours, slices were stored at-80~C for Western-blot a- nalysis. Results In contrast to the low level of cx32 immunohistochemistry expresses in the control hippocampal tissue, the strongly positive intensity of cx32 and cx43 in 14 cases of TLE group were found. Cx32 was expressed at a significantly higher level in the epileptic patients and at those after VPA treatment compared with that of the controls and those after Carb treatment by Western-blot（ P 〈 0.05 ） ,9 hours after Carb treatment ,levels of Cx32 was decreased ,respectively,there was no change after VPA treatment. However, There was no difference of Cx32 expression in the control hippocampal tissue and those after Carb treatment （P 〉 0.05 ）. Conclusion The increase in Cx32 expression in hippocampi from epileptic per- sons could play a role in the development of seizures in patients with temporal seizures. Additional studies must be comple- ted to understand this mechanism better.

关 键 词：缝隙连接 颞叶癫痫 海马 生胃酮 丙戊酸钠Cx32 

分 类 号：R742.1[医药卫生—神经病学与精神病学]