VDAC1在富氢液减轻大鼠全脑缺血再灌注损伤中的作用  

Role of voltage-dependent anion channel 1 in attenuation of global cerebral ischemia-reperfusion injury by hydrogen-rich saline in rats

作　　者：戴琼艳 龙云[2] 赵琳[2] 张露[1] 高宇[2] 吴淋 陈兴东[2] 段满林[2] 徐建国[2] 

机构地区：[1]徐州医学院江苏省麻醉学重点实验室,221000 [2]南京大学医学院南京军区南京总医院麻醉科,210002

出　　处：《中华麻醉学杂志》2014年第12期1481-1484,共4页Chinese Journal of Anesthesiology

摘　　要：目的 评价电压依赖性阴离子通道1（VDAC1）在富氢液减轻大鼠全脑缺血再灌注损伤中的作用.方法 雄性SD大鼠60只,体重280 - 320 g,采用随机数字表法,将其分为6组（n=10）：假手术组（S组）、缺血再灌注组（I/R组）、富氢液组（H组）、二甲基亚砜溶剂对照组（DMSO组）、VDAC1阻断剂4＇-二异硫氰基芪-2,2＇-二磺酸组（DIDS组）和富氢液＋DIDS组（HD组）.采用经食管心脏起搏诱发室颤后心肺复苏建立大鼠全脑缺血再灌注损伤模型.H组和HD组于再灌注即刻和6h时腹腔注射富氢液5 ml/kg,I/R组腹腔注射等容量生理盐水;DIDS组和HD组诱发室颤前经15 min股静脉输注DIDS 12 mg/kg,次日9时腹腔注射DIDS 12 mg/kg;DMSO组诱发室颤前经15 min股静脉输注DIDS的溶剂DMSO 12 mg/kg,次日9时腹腔注射DMSO 12 mg/kg.于再灌注24h时行神经功能评分,然后处死大鼠,取海马组织,计数正常椎体细胞,采用Western blot法测定VDAC1蛋白、细胞色素c和活化的caspase-3表达水平,采用荧光探针法测定活性氧（ROS）的含量.结果 与S组比较,其余各组神经功能评分和正常锥体细胞计数降低,海马组织VDAC1蛋白、细胞色素c和活化的caspase-3表达上调,ROS含量升高（P＜0.05）;与I/R组比较,H组、DIDS组和HD组神经功能评分和正常锥体细胞计数升高,海马组织VDAC1蛋白、细胞色素c和活化的caspase-3表达下调,ROS含量降低（P＜0.05）,DMSO组上述各指标差异无统计学意义（P＞0.05）;与H组比较,HD组神经功能评分和正常锥体细胞计数升高,海马组织VDAC1蛋白、细胞色素c和活化的caspase-3表达下调,ROS含量降低（P＜0.05）.结论 富氢液通过抑制VDAC1的开放减轻大鼠全脑缺血再灌注损伤.Objective To evaluate the role of voltnge-dependent anion channel 1 （VDAC1） in attenuation of global cerebral ischemia-reperfusion （I/R） injury by hydrogen-rich saline in rats.Methods Sixty male Sprague Dawley rats,weighing 280-320 g,were randomly divided into 6 groups （n =10 each） using a random number table：sham operation group （group S）,I/R group,hydrogen-rich saline group （group H）,vehicle dimethyl sulfoxide （DMSO） group,VDAC1 blocker 4,4＇-diisothiocyanostilbene-2,2＇-disulfonic acid （DIDS） group （group DIDS）,and hydrogen-rich saline ＋ DIDS group （group HD）.Ventricular fibrillation （VF） was induced by transoesophageal cardiac pacing and cardiopulmonary resuscitation was initiated 4 min after onset of VF to establish the model of global cerebral I/R injury.In H and HD groups,hydrogen-rich saline 5 ml/mg was injected intraperitoneally at 0 and 6 h after reperfusion,while the equal volume of normal saline was injected instead of hydrogen-rich saline in group I/R.In DIDS and HD groups,DIDS 12 mg/kg was infused over 15 min via the femoral vein prior to induction of VF,and DIDS 12 mg/kg was injected intraperitoneally at 9：00 am on the next day.In DMSO group,DMSO 12 mg/kg was infused over 15 min viathe femoral veprior to induction of VF,and DMSO 12 mg/kg was injected intraperitoneally at 9：00 am on the next day.After the neurological score was evaluated at 24 h of reperfusion,the rats were sacrificed.Hippocampi were immediately removed for determination of the number of normal pyramidal neurons,expression of VDAC1 protein,cytochrome c and activated caspase-3 （by Western blot）,and content of reactive oxygen species （ROS） （by using fluorescence probe）.Results Compared with group S,the neurological scores and the number of normal pyramidal neurons were significantly decreased,the expression of VDAC1 protein,cytochrome c and activated caspase-3 was up-regulated,and the content of ROS was increased in the other groups.Compared with group I/R,the neurologicals

关 键 词：电压依赖性阴离子通道1 氢 再灌注损伤 脑 

分 类 号：R743.3[医药卫生—神经病学与精神病学]