Nrf2基因敲除对小鼠蛛网膜下腔出血后脑损伤的作用  

Nrf2 knockout and brain injury following subarachnoid hemorrhage in mice

作　　者：李桃[1] 王汉东[1] 丁宇[1] 何进[1] 丁可[1] 陆新宇[1] 徐建国[1] 

机构地区：[1]南京大学医学院附属金陵医院(南京军区南京总医院)神经外科,南京医学硕士210002

出　　处：《医学研究生学报》2014年第11期1128-1132,共5页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81070974)

摘　　要：目的蛛网膜下腔出血(subarachnoid hemorrhage,SAH)是一种致死率较高的危重疾病,文中研究氧化应激调节因子Nrf2在SAH后脑损伤作用及机制。方法实验选取雄性ICR野生型(wild type,WT)小鼠及来源于ICR的Nrf2基因敲除(knockout,KO)小鼠,采用视交叉自体血注射建立小鼠SAH模型,实验动物分为WT假手术组、KO假手术组、WT SAH组和KO SAH组4个组,检测SAH后24 h氧化应激产物丙二醛(malondialdehyde,MDA)及GSH/GSSG,炎症因子TNF-α和IL-1β,脑组织含水量和伊文思蓝含量,TUNEL和尼氏染色,活动评分及大脑前和大脑中动脉血管痉挛情况。结果与假手术组比较,SAH组MDA、TNF-α、IL-1β表达量上升,而GSH/GSSG下降(P<0.01);与WT SAH组比较,MDA、TNF-α、IL-1β表达量上升(P<0.05),而GSH/GSSG下降(P<0.05)。SAH组前脑脑组织含水量、伊文思蓝含量较假手术组增加(P<0.01),与WT SAH组比较,KO SAH组脑组织含水量、伊文思蓝含量均升高[(0.808±0.004)vs(0.819±0.004)、(7.230±1.192)μg/g vs(11.628±1.040)μg/g,P<0.05]。SAH后24 h,与假手术组比较,SAH组神经细胞凋亡率上升(P<0.01),而神经元数量、ACA比值、血管半径/壁厚值、活动评分下降(P<0.01),与WT SAH组比较,KO SAH组细胞凋亡率上升[(23.733±8.204)%vs(36.267±10.612)%],而神经元数、ACA比值、血管半径/壁厚值、活动评分下降[(70.833±8.750)vs(51.767±13.006),(8.024±2.780)vs(6.861±2.702),(6.337±3.993)vs(5.107±3.805),(1.967±0.928)vs(1.433±0.679),P<0.05]。结论 Nrf2 KO加重了SAH后氧化应激和炎性反应,从而导致了SAH继发性脑损伤加重。Nrf2对SAH后继发性脑损伤具有保护作用。Objective Subarachnoid hemorrhage （ SAH） is a devastating disease with a high mortality.This study was to in-vestigate the effect of Nrf2 on secondary brain injury following SAH and its action mechanism in mice. Methods SAH models were established in wild-type （ WT） and Nrf2 knockout （ KO） ICR male mice by injecting fresh blood drawn from the femoral artery into the pre-chiasmatic cistern.The animals were divided into four groups, WT sham, WT SAH, KO sham, and KO SAH.At 24 hours after modeling, the expression levels of malondialdehyde （ MDA） , GSH/GSSG, TNF-αand IL-1β, the volume of brain water, and content of Evans blue were measured, the activity scores obtained, and cerebral vasospasm of the anterior and middle cerebral arteries （ ACA and MCA） detected. Results At 24 hours, the expressions of MDA, TNF-α, and IL-1βwere （3.299 ±0.335）, （1.187 ± 0.436）, and （59.330 ±21.787） mg/g in the WT sham group, （4.339 ±0.328）, （2.432 ±0.434）, and （121.584 ±21.675） mg/g in the WT SAH group, （3.488 ±0.634）, （1.170 ±0.312）, and （58.497 ±15.608） mg/g in the KO sham group, and （5.335 ±0.499）, （3.132 ±0.548）, and （171.117 ±50.479） mg/g in the KO SAH group, markedly increased in the SAH groups as compared with the sham controls （P〈0.05）, while the GSH/GSSG levels were significantly higher in the former two groups than in the latter （0.553 ±0.100 and 0.375 ±0.068 vs 0.714 ±0.091, 0.761 ±0.114, P〈0.01）.The contents of brain water and Evans blue were （0.784 ±0.005） and （7.055 ±1.046） μg/g in the WT sham group, （0.808 ±0.004） and （7.230 ±1.192） μg/g in the WT SAH group, （0.784 ±0.004） and （9.620 ±1.290） μg/g in the KO sham group, and （0.819 ±0.004） and （11.628 ±1.040）μg/g in the KO SAH group, remarkably increased in the SAH groups in comparison with the sham groups （P〈0.05）.The apoptosis rate 8.916 and 82.100 ±6.870 vs 70.833 ±8.750 and 51.767 ±13.006）, ACA radius/wall thickness val

关 键 词：核因子E2相关因子2 蛛网膜下腔出血 氧化应激 炎症因子 早期脑损伤 脑血管痉挛 NF-E2-related factor 2 

分 类 号：R442.7[医药卫生—诊断学]