浅低温联合线粒体分裂抑制剂减轻全脑缺血-再灌注后线粒体损伤  

Mild hypothermia combined with mitochondrial division inhibitor alleviates mitochondrial damage after global cerebral ischemia-reperfusion

作　　者：范婧婧[1] 惠康丽[2] 徐苗苗[2] 仲浩 蔡慎权 操良斌 段满林[2] 徐建国[2] 

机构地区：[1]南京大学医学院,210002 [2]南京军区南京总医院 [3]徐州医科大学

出　　处：《临床麻醉学杂志》2017年第7期697-700,共4页Journal of Clinical Anesthesiology

基　　金：国家自然科学基金(No.81671884)

摘　　要：目的观察浅低温联合线粒体分裂抑制剂(mitochondrial division inhibitor 1,Mdivi-1)对大鼠海马缺血-再灌注(ischemia-reperfusion,IR)损伤后线粒体的影响。方法健康清洁级雄性SD大鼠40只,体重280~320g,采用随机数字表法分为五组:假手术组(Sham组)、全脑IR组(IR组)、浅低温组(H组)、线粒体分裂抑制剂组(M组)和浅低温联合线粒体分裂抑制剂组(HM组),每组8只。采用经食管心脏起搏诱发心跳骤停+心肺复苏方法制备大鼠全脑IR损伤模型,即缺血4min,再灌注6h。Sham组不经历心跳骤停和心肺复苏,其余操作与IR组一致;H组和HM组于再灌注即刻体表降温,15min之内将直肠温度降至32~34℃,自制变温箱维持6h,其余组维持正常体温;M组和HM组于再灌注即刻静脉注射Mdivi-1(1.2 mg/kg),其余组注射等容积二甲基亚砜(DMSO)。于再灌注6h处死大鼠取双侧海马,采用Western blot法检测线粒体发动相关蛋白-1(dynamin-related protein 1,Drp1)和细胞色素C(cytochrome Cyt-C)蛋白含量,电镜下观察海马区椎体细胞中线粒体结构形态。结果 IR组、H组、M组和HM组Drp1和Cyt-C蛋白含量明显高于Sham组(P<0.05);H组、M组和HM组Drp1和Cyt-C蛋白含量明显低于IR组(P<0.05);HM组Drp1和Cyt-C蛋白含量明显低于H组和M组(P<0.05),H组和M组Cyt-C蛋白含量差异无统计学意义。Sham组线粒体呈短棒状,结构清晰完整,其他各组中可见不同程度的线粒体分裂,膜间隙肿胀,基质包含物沉积,空泡形成,嵴模糊或消失,HM组病变较轻。结论浅低温联合线粒体分裂抑制剂可以减轻大鼠全脑缺血-再灌注后线粒体损伤,联合效果优于单独应用。Objective To investigate the effect of mild hypothermia combined with mitochondrial divison inhibitor 1 in mitochondrial after cerebral ischemia-reperfusion （IR）.Methods Fourty male healthy Sprague-Dawley （SD） rats, weighing 280-320 g, were randomly divided into 5 groups （n=8 each）： group Sham, group IR, hypothermia group （group H）, Mdivi-1 group （group M） and hypothermia＋Mdivi-1 group （group HM）.Animal models of global cerebral IR were established by transoesophageal cardiac pacing inducing cardiac arrest followed by cardiopulmonary resuscitation （ischemia 4 min and reperfusion 6 h）.The group Sham was similarly treated to group IR except the cardiac arrest and cardiopulmonary resuscitation.In groups H and HM, the core temperature was cooled down to 32-34℃ within 15 min starting from the beginning of reperfusion, and maintained for 6 h.In the other groups, the core temperature was maintained at the normal temperature.In groups M and HM, the animals were given Mdivi-1 （1.2 mg/kg） intravenously at the beginning of the reperfusion and the other groups were given the same volumn of dimethylsnlfone （DMSO）.After 6 h of reperfusion, the rats were sacrificed, and bilateral hippocampi were immediately removed for determination the protein level of dynamin-related proten 1 （Drp1） and cytochrome C （Cyt-C） expression by Western blot and obsevation of the mitochondrial structure of pyramidal cell in hippocampal CA1 under electronic microscope.Results Compared with group Sham, the expression of Drp1 and Cyt-C was up-regulated in groups IR, H, M and HM （P〈0.05）.Compared with group IR, the expression of Drp1 and Cyt-C was down-regulated in groups H, M and HM （P〈0.05）.Compared with groups H and M, the expression of Drp1 and Cyt-C was down-regulated in group HM （P〈0.05）.There was no significant difference in the expression of Drp1 and Cyt-C between groups H and M.The mitochondria were rod-shaped with clear and sound structure in group Sham, while mitochondria show

关 键 词：浅低温 线粒体分裂抑制剂 全脑缺血-再灌注 发动相关蛋白-1 细胞色素C 

分 类 号：R61[医药卫生—外科学]