摘要
目的观察腺苷激酶(ADK)在难治性癫痫患者海马内的表达,进而探讨ADK在难治性癫痫发病机制中的作用。方法收集哈尔滨医科大学附属第一医院因难治性癫痫手术切除海马组织的患者13例,作为癫痫组;同时收集颅脑外伤行清创术患者(相对正常的颞叶脑组织)4例(既往无癫痫发作史),该4例患者为对照组。利用双荧光免疫组织化学、实时定量PCR(RT-Real-time PCR)、蛋白免疫印迹等方法从组织水平、基因水平、蛋白水平检测两组标本ADK的表达情况。结果在人脑中,ADK主要在星形胶质细胞核中表达。通过组织学观察,ADK在正常脑组织内有微弱表达,而在难治性癫痫患者脑组织中胶质细胞明显增生,同时有ADK的过量表达。癫痫组ADK阳性胶质细胞百分比为(53.90±17.59)%,对照组为(23.82±4.18)%,差异有统计学意义(P<0.01)。从基因水平,利用RT-Real-time PCR法发现癫痫组ADK mRNA高于对照组表达量,其2^(-△△Cp)为13.36,表达量是对照组的13.36倍。在蛋白水平,利用蛋白免疫印迹方法发现癫痫组ADK蛋白表达高于对照组(P<0.01)。结论ADK在人的正常脑组织星形胶质细胞核内有微弱表达。而在难治性癫痫患者脑组织中,星形胶质细胞显著增生并且有ADK的过量表达。ADK在人类难治性癫痫的发生发展过程中可能起到了重要作用。
Objective To observe the expression of adenosine kinase(ADK)in the hippocampus of patients with refractory epilepsy,and to explore the role of ADK in the pathogenesis of refractory epilepsy.Methods Thirteen patients with intractable epilepsy who underwent surgical resection of hippocampal tissue at the First Affiliated Hospital of Harbin Medical University were collected as the epilepsy group;At the same time,4 cases of relatively normal temporal lobe brain tissue from patients with traumatic brain injury undergoing debridement surgery(without previous history of epileptic seizures)were collected,and these 4 patients served as the control group.The expression of ADK in two groups of specimens was detected at the tissue,gene,and protein levels using methods such as dual fluorescence immunohistochemistry,real-time quantitative polymerase chain reaction(RT Real time PCR),and Western blotting.Results In the human brain,ADK was mainly expressed in the nucleus of astrocytes.Through histological observation,ADK was weakly expressed in normal brain tissue,while there is significant proliferation of glial cells and excessive expression of ADK in the brain tissue of patients with refractory epilepsy.The percentage of ADK positive glial cells in the epilepsy group was(53.90±17.59)%,and the control group was(23.82±4.18)%,with a statistically significant difference(P<0.01).At the genetic level,using RT Real time PCR,it was found that the expression level of ADK mRNA in the epilepsy group was higher than that in the control group,with a 2^(-△△Cp) of 13.36,which was 13.36 times higher than that in the control group.At the protein level,the expression of ADK protein in the epilepsy group was found to be higher than that in the control group using protein immunoblotting(P<0.01).Conclusions ADK is weakly expressed in the nucleus of astrocytes in normal human brain tissue.In the brain tissue of patients with refractory epilepsy,astrocytes significantly proliferate and there is excessive expression of ADK.ADK may play an impor
作者
赫丹丹
兰春伟
寇雨霏
刘涛
王清勇
段淑荣
He Dandan;Lan Chunwei;Kou Yufei;Liu Tao;Wang Qingyong;Duan Shurong(Department of Neurology,Shenzhen Hospital,University of Chinese Academy of Sciences,Shenzhen 518107,China;Department of Neurology,the First Affiliated Hospital of Harbin Medical University,Harbin 150001,China)
出处
《中国医师杂志》
CAS
2023年第9期1355-1358,共4页
Journal of Chinese Physician
基金
深圳市光明区软科学研究项目(2021R01017)。
关键词
腺苷激酶
难治性癫痫
海马
Adenosine kinase
Intractable epilepsy
Hippocampus
