酒精依赖因素对大鼠脊髓神经元K＋-Cl-共转运体2表达的影响  

Effects of alcohol dependence on expression of spinal neuronal K＋-Cl-cotransporter 2 in rats

作　　者：王红柏[1] 王海云[1] 王国林[1] 刘书颖 朱爱[1] 

机构地区：[1]天津医科大学总医院麻醉科天津市麻醉学研究所,300052

出　　处：《中华麻醉学杂志》2015年第4期466-469,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金（81071059,81100984,81371245）;天津市卫生局科技基金（09KZ106）

摘　　要：目的 探讨酒精依赖因素对大鼠脊髓神经元K＋-Cl共转运体2（KCC2）表达的影响.方法 健康雄性SD大鼠48只,体重200～ 250 g,采用随机数字表法,将其分为2组（n=24）：对照组（C组）和酒精依赖组（AD组）.采用经口插管灌胃的方法给予酒精.第1、2、3周酒精浓度分别为5％、10％、20％,第4周及以后浓度为35％,灌胃量为10 ml·kg-1·d-1,总时程为8周.C组采用同样方法经口灌注不含酒精的饮用水10 ml·kg-1·d-1.2组大鼠均于最后一次灌胃前行高架十字迷宫实验.灌胃结束后建立大鼠切口痛模型.于术后2、6、24和48 h时测定机械痛阈和热痛阈.于术后48 h时处死大鼠,取脊髓组织,采用免疫荧光法和Western blot法测定脊髓KCC2的表达水平.结果 与C组比较采,AD组开放臂进入次数减少,开放臂停留时间缩短,闭合臂进入次数增多,闭合臂停留时间延长,术后各时点机械痛阈和热痛阈降低,脊髓KCC2表达下调（P＜0.05）.结论 酒精依赖大鼠痛觉过敏的机制与脊髓神经元KCC2表达下调有关.Objective To evaluate the effects of alcohol dependence （AD） on the expression of spinal neuronal K＋-Cl cotransporter 2 （KCC2） in rats.Methods Forty-eight healthy male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 2 groups （n =24 each） using a random number table：control group （group C） and group AD.An orogastric tube was inserted and alcohol was administered through the tube into the stomach to establish the model of AD.The concentration of ethanol was 5％,10％ and 20％ at 1st,2nd and 3rd weeks,respectively,and the concentration of ethanol was 35％ at 4th week and later.Alcohol was given at 10 ml · kg-1 · d-1,lasting for 8 weeks.The rats received drinking water containing no ethanol at 10 ml · kg-1 · d-1 instead of alcohol in group C.All the rats were allowed ad libitum access to food and water.Before the last administration,an elevated plus-maze test was performed for all the rats to observe their state of anxiety,which was used to evaluate the success of AD model.At the end of the last administration,the model of incisional pain was established.A 1-cm longitudinal incision was made through skin,fascia and muscle of the plantar aspect of the hindpaw in sevoflurane-anesthetized rats.At 2,6,24 and 48 h after operation,the mechanical and thermal paw withdrawal thresholds were measured.At 48 h after operation,the lumbar segment of the spinal cord was removed for determination of the expression of KCC2 by using immunofluorescence and Western blot.Results Compared with group C,the number of open arm entries was significantly reduced,the time spent on the open arms was shortened,the number of closed arm entries was increased,the time spent on the closed arms was prolonged,the mechanical and thermal paw withdrawal thresholds were decreased,and the expression of KCC2 was down-regulated in group AD.Conclusion Down-regulated expression of spinal neuronal KCC2 is involved in the mechanism of hyperalgesia in rats with AD.

关 键 词：酒精中毒 协同转运子 痛觉过敏 脊髓 神经元 

分 类 号：R614[医药卫生—麻醉学]