腺苷酸激酶4在脑缺血早期神经损伤中的作用研究  

Study on the Role of Adenylate Kinase 4 in Nerve Injury in the Early Stage of Cerebral Ischemia

作　　者：钟云雪 谢聪 胡凌慧 贾冰冰 廖子君 张圆[3] 刘文兰[3] ZHONG Yunxue;XIE Cong;HU Linghui;JIA Bingbing;LIAO Zijun;ZHANG Yuan;LIU Wenlan(Hengyang Medicine School,University of South China,Hunan Hengyang 421001;Graduate School of Guangzhou Medical University,Guangdong Guangzhou 511436;Shenzhen Second People's Hospital,The First Affiliated Hospital of Shenzhen University,Guangdong Shenzhen 518035;Graduate Guangxi University of Chinese Medicine,Guangxi Nanning 530200)

机构地区：[1]南华大学衡阳医学院,湖南衡阳421001 [2]广州医科大学研究生院,广东广州511436 [3]深圳市第二人民医院深圳大学第一附属医院,广东深圳518035 [4]广西中医药大学研究生院,广西南宁530200

出　　处：《深圳中西医结合杂志》2024年第4期1-5,I0006,共6页Shenzhen Journal of Integrated Traditional Chinese and Western Medicine

基　　金：广东省自然科学基金面上项目(2022A1515010466);深圳市科技计划项目(JCYJ20230807115310022)。

摘　　要：目的:脑供血不足导致的神经元损伤是缺血性脑卒中神经功能障碍的直接原因,线粒体功能异常与神经元损伤密切相关,腺苷酸激酶4(AK4)是能量代谢关键因子,本研究旨在探讨AK4在脑缺血早期神经损伤中的作用。方法:研究样本为60只雄性C57BL/6J小鼠,3~4周龄,体质量18~20 g,构建神经元特异性过表达AK4腺相关病毒(AAV)-空载(AAV-Ctrl组)以及AAV-AK4过表达(AAV-AK4组)小鼠。通过建立小鼠大脑中动脉阻塞(MCAO)模型模拟体内脑缺血,分别在缺血1 h、2 h和3 h后采用免疫印迹法测定AK4的蛋白表达水平;在单纯缺血3 h后,分别采用2,3,5-氯化三苯基四氮唑(TTC)染色法检测缺血脑组织损伤程度,末端脱氧核苷酸转移酶介导的dUTP缺口末端标记测定法(TUNEL)法检测神经元凋亡情况。结果:实验结果表明,MCAO 2 h和MCAO 3 h时梗死侧(I)相较于非梗死侧(NI)的AK4蛋白表达开始下调,其中MCAO 3 h最明显,差异均具有统计学意义(P <0.05)。与AAV-Ctrl组比较,MCAO 3 h后,AAV-AK4组小鼠的脑梗死体积明显缩小,且AAV-AK4组比AAV-Ctrl组小鼠的神经元凋亡数量明显减少,差异均具有统计学意义(P <0.05)。结论:脑缺血早期神经元过表达AK4可明显减少脑梗死体积,抑制神经元凋亡,发挥神经保护作用,AK4可能是缺血早期神经元损伤干预的新靶点。Objective Neuronal damage caused by insufficient cerebral blood supply is the direct cause of neurological dysfunction in ischemic stroke.Mitochondrial dysfunction is closely related to neuronal damage.Adenylate kinase 4(AK4)is a key factor in energy metabolism.The purpose of this study is to explore the role of AK4 in early nerve injury after cerebral ischemia.Methods The study sample consisted of 60 male C57BL/6J mice,3 to 4 weeks of age and 18 to 20 g body mass,constructed neuron-specific overexpressing AK4 adeno-associated virus(AAV)-unloaded(AAV-Ctrl group)and AAV-AK4 overexpressing mice(AAV-AK4 group).A mouse middle cerebral artery occlusion(MCAO)model was established to simulate cerebral ischemia in vivo,and the protein expression level of AK4 was determined by western blot after 1 h,2 h and 3 h of ischemia,respectively.After 3 h of pure ischemia,2,3,5-triphenyltetrazolium chloride(TTC)staining was used to detect the degree of ischemic brain tissue injury,and TUNEL method was used to detect neuronal apoptosis.Results The experimental results showed that AK4 protein expression on infarct side(I)began to be down-regulated compared with non-infarct side(NI)at MCAO 2 h and MCAO 3 h,and MCAO 3 h was the most significant,with statistical significance(P<0.05).Compared with AAV-Ctrl group,after 3 h of MCAO,the volume of cerebral infarction in AAV-AK4 group was significantly reduced,and the number of neuronal apoptosis in AAV-AK4 group was significantly reduced compared with AAV-Ctrl group,with statistical significances(P<0.05).Conclusion Overexpression of AK4 in neurons in the early stage of cerebral ischemia can significantly reduce the volume of cerebral infarction inhibit neuronal apoptosis and play a neuroprotective effect. AK4 may be a new target for intervention of neuronal injury in the early stage of cerebral ischemia.

关 键 词：腺苷酸激酶4 缺血性脑卒中 动物实验 小鼠 

分 类 号：R741[医药卫生—神经病学与精神病学] R-332[医药卫生—临床医学]