脑缺血再灌注小鼠线粒体DNA拷贝数与神经功能损伤相关性研究  

Association between mitochondrial DNA copy number and neurological severity in cerebral ischemia mice

作　　者：付浩[1,2] 胡文静 王志宏[1] 陈孝储[1] 涂悦[1,2] 程世翔[1,2] 孙圣凯[1] 

机构地区：[1]武警后勤学院附属医院,天津300162 [2]天津市神经创伤修复重点实验室/武警部队脑创伤与神经疾病研究所,天津300162 [3]武警重庆总队第七支队卫生队,重庆401347

出　　处：《现代中西医结合杂志》2017年第1期4-6,13,共4页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：天津市卫生局科技基金项目(2015kz126);天津市卫生计生委中医中西医结合科研课题(2015155);天津市重点科研平台开放基金项目(WYKFZ201602);武警后勤学院科学研究项目(WHJ2015013);武警后勤学院附属医院种子基金项目(FYM201517;FYQ201560)

摘　　要：目的探讨小鼠脑缺血再灌注后外周血白细胞线粒体DNA拷贝数变化及其与神经功能损伤程度的相关性。方法将40只C57BL/6小鼠随机分为假手术组、脑缺血再灌注24 h组、脑缺血再灌注48 h组、脑缺血再灌注72 h组,每组10只。线栓法构建大脑中动脉闭塞(MCAO)模型并在1 h后行再灌注。脑缺血再灌注各组分别于再灌注相应时间点进行神经功能损伤评分(mNSS)评价,随后立即取血并提取基因组DNA,real-time PCR法检测相对线粒体拷贝数。结果脑缺血再灌注24 h组、脑缺血再灌注48 h组线粒体DNA拷贝数均明显高于假手术组(P均<0.05),线粒体DNA拷贝数与mNSS评分呈正相关(r=0.639,P<0.05;r=0.667,P<0.05);脑缺血再灌注72 h组线粒体DNA拷贝数与假手术组比较差异无统计学意义(P>0.05),线粒体DNA拷贝数与mNSS评分不具有相关性(r=0.420,P>0.05)。结论外周血白细胞线粒体DNA拷贝数在小鼠脑缺血再灌注24 h和48 h后显著增加,且拷贝数越高,神经功能损伤程度越严重。Objective It is to explore the changes of peripheral leukocytes mitochondrial DNA copy number in cerebral is- ehemia reperfusion mice and its correlation with neurological severity. Methods Forty male C57BL/6 mice were randomly di- vided into middle cerebral artery occlusion （MCAO）group （30 mice divided into 3 group equally） and shame group （10 mice）. The MCAO models were conducted by the intraluminal filament technics and reperfusion was made 1 h later. At differ- ent Lime points after operation （24 h, 48 h, 72 h） , neurological function was evaluated according to modified Neurological Se- verity Score （mNSS） , genomic DNA was extracted from blood. Mitochondrial DNA copy number was measured by real-time PCR. Results At 24 h and 48 h after the operation, the mitochondrial DNA copy number was significantly increased （P 〈 O. 05 ） , and was positively associated with mNSS （ r = 0. 639,P 〈 0.05 and r = 0. 667, P 〈 0.05 ）. At 72 h after operation, the mitochondrial DNA copy number was significantly decreased when compared with the 24 h and the 48 h group, and no sig- nificant difference with shame group was found, the copy number was not associated with mNSS. Conclusion Mitochondrial DNA copy number was significantly increased at 24 h and 48 h after MCAO operation, and the more mitochondrial DNA copy numbers are, the weaker neurological function is.

关 键 词：脑缺血再灌注 线粒体 神经功能损伤评分 氧化应激 大脑中动脉闭塞 

分 类 号：R-332[医药卫生]