乙酰唑胺对戊四氮致慢性癫痫大鼠海马水通道蛋白4表达的影响  

Effect of Acetazolamide on the Expression of Aquaporin-4 in Hippocampus of Rats with PTZ-induced Chronic Epilepsy

作　　者：陈丽[1] 余涵[1] 周玉波[1] 马猛[1] 王林杰[1] 胡景鑫[1] 雷水生[2] 朱晓琴[1] 

机构地区：[1]广州医科大学基础学院,广州511436 [2]广州医科大学附属第五医院皮肤美容科,广州510700

出　　处：《华中科技大学学报（医学版）》2015年第5期510-514,519,共6页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：广州市科技计划项目(No.2013J4100103);广州市属高校科研计划项目(No.2012C128;2012D001);广东省省级科技计划项目(No.2013B021800191;2014A020212512)

摘　　要：目的观察乙酰唑胺(acetazolamide,AZA)对戊四氮(pentylenetetrazole,PTZ)致慢性癫痫大鼠发作及海马水通道蛋白4(aquaporin-4,AQP4)表达的影响,探讨其抗癫痫的可能机制。方法将实验大鼠随机分为4组:对照组(每日腹腔注射生理盐水)、致痫组[每日腹腔注射亚惊厥剂量PTZ 35mg/(kg·d)建立慢性癫痫大鼠模型]、乙酰唑胺干预组[每日先腹腔注射AZA 35mg/(kg·d)预处理,30min后再腹腔注射亚惊厥剂量PTZ]和乙酰唑胺对照组[每日单纯腹腔注射AZA]。各组采用以上注射方式连续给药28d,每次注射后1h内记录各组大鼠癫痫发作级别,统计各组每天平均发作级别,观察乙酰唑胺对大鼠慢性癫痫发作的影响;最后1次给药1d后,脑电图记录各组大鼠脑电的改变,免疫组化观察AQP4在海马内分布,实时荧光定量PCR(RT-qPCR)和Western blot检测AQP4mRNA和蛋白的表达。结果对照组和乙酰唑胺对照组无明显癫痫发作,乙酰唑胺干预组潜伏期为(23.5±2.4)d,而致痫组潜伏期为(18.2±1.7)d,乙酰唑胺干预可明显延长慢性癫痫大鼠点燃的潜伏期(P<0.05);脑电图结果对照组和乙酰唑胺对照组无明显痫样波,致痫组记录到棘波、尖波等痫样波,而乙酰唑胺干预组痫样波明显减弱;免疫组化结果显示:AQP4主要分布在海马CA3区血管周围;RT-qPCR和Western blot结果显示慢性致痫组AQP4 mRNA及蛋白表达明显增加(P<0.05或P<0.01),乙酰唑胺干预组AQP4表达降低但仍高于对照组(P<0.05),乙酰唑胺对照组AQP4表达较对照组明显降低(P<0.05)。结论乙酰唑胺能够抑制癫痫的发作,延长慢性点燃潜伏期,并且抑制AQP4的表达,其作用机制可能是与乙酰唑胺抑制AQP4表达,改善慢性癫痫状态下脑内水和离子平衡有关。Objective To observe the effect of acetazolamide（AZA）on pentylenetetrazole（PTZ）-induced chronic epileptic seizures in rats and aquaporin-4（AQP4）expression in hippocampus tissue of rats,and investigate the possible antiepileptic mechanism of AZA.Methods The experimental rats were randomly divided into four groups：control group,in which the rats were daily intraperitoneally injected with normal saline;epileptic group,in which the rats were daily administered a subconvulsive dose of PTZ（35mg/kg）;AZA＋PTZ group,in which the rats were daily treated with AZA（35mg/kg）30min before PTZ injection;AZA group,in which the rats were intraperitoneally injected with AZA（35mg/kg）each day.All groups were administered for 28 consecutive days.The seizure severity of rats within one hour after each injection was recorded and the average daily seizure severity was obtained in each group to observe the effect of AZA on chronic epileptic seizures.One day after the last administration,the electroencephalogram changes of rats were recorded.Immunohistochemistry was used to detect the distribution of AQP4 in rat hippocampus.Real-time quantitative PCR（RT-qPCR）and Western blot were used to detect the expression of AQP4.Results Behavioral observation showed that rats in control group and AZA group had no obvious epileptic seizure.The average latent period was significantly increased in AZA＋PTZ group（23.5±2.4）d compared with that in the PTZ group（18.2±1.7）d（P〈0.05）.AZA intervention could significantly prolong the latent period of chronic epilepsy.EEG showed no significant epileptic waves in the control group and the AZA group,but spike wave and sharp wave in the epileptic group.In the AZA＋PTZ group,the number of epileptic waves was conspicuously reduced.Immunohistochemistry showed AQP4 was primarily localized surrounding brain capillaries in hippocampus CA3 region.The results of RT-qPCR and Western blot showed that the ex-pression level of AQP4 mRNA and protein in hippocampus of PTZ group was i

关 键 词：慢性癫痫 水通道蛋白4 乙酰唑胺 戊四氮 

分 类 号：R742.1[医药卫生—神经病学与精神病学]