Neuritin对大鼠蛛网膜下腔出血后早期脑损伤影响的研究  

Effects of Neuritin in Early Brain Injury After Subarachnoid Hemorrhage of Rats

作　　者：李晓天[1,2] 赵冬[1] 代林志[1] 朱立仓[1] 田卫东[1] 唐仕军[1] 朱文学[1] 杨鹏[1] 王业忠[1] 

机构地区：[1]石河子大学医学院第一附属医院神经外科,新疆石河子市832000 [2]潍坊市人民医院

出　　处：《中国全科医学》2016年第24期2908-2914,共7页Chinese General Practice

基　　金：国家自然科学资金资助项目(81360185)

摘　　要：目的探究Neuritin对大鼠蛛网膜下腔出血(SAH)后早期脑损伤过程中内质网应激因子GRP78及血-脑脊液屏障通透性的影响。方法 2014年4月—2015年9月选取成年健康雄性SD大鼠144只,按照随机数字表法分为对照组、假手术组(Sham组)、SAH组、Neuritin组,各36只。再依据简单随机抽样法分别于3、6、12、24、48、72 h抽取每组6只大鼠处死,Neuritin组于处死前1 h采用立体定向技术向侧脑室注射Neuritin。建模后6、12、24、48、72 h处死大鼠前,记录各组大鼠Garcia神经功能评分;建模后3、6、12、24、48、72 h处死大鼠后,采用HE染色观察SAH建立情况,侧脑室微量注射亚甲蓝鉴定Neuritin注射情况;采用Western blotting法检测大鼠大脑皮质内质网应激因子GRP78表达水平;采用甲酰胺浸泡法检测大鼠脑内伊文蓝(EB)含量以评价血-脑脊液屏障通透性。结果 SAH组、Neuritin组建模后6、12、24、48、72 h大鼠Garcia神经功能评分均低于对照组和Sham组(P<0.05);Neuritin组建模后6、12、24、48、72 h大鼠Garcia神经功能评分均高于SAH组(P<0.05)。SAH组大鼠大脑表面弥散分布积血,其中在willis环、小脑延髓池及脑干腹侧有大量积血存在。Neuritin组大脑腹面有亚甲蓝循环存在,大脑背面无亚甲蓝,Neuritin侧脑室注射成功。SAH组建模后3、6、12、24、48、72 h GRP78表达水平高于对照组、Sham组(P<0.05);Neuritin组建模后6、12、24、48、72 h GRP78表达水平低于SAH组(P<0.05)。SAH组大鼠建模后3、6、12、24、48、72 h脑组织EB含量均高于对照组、Sham组(P<0.05);Neuritin组大鼠建模后3、12 h脑组织EB含量高于SAH组,建模后6、48、72 h脑组织EB含量低于SAH组(P<0.05)。结论 SAH后早期脑损伤过程中存在内质网应激反应,神经营养因子Neuritin可显著改变内质网应激因子GRP78表达水平,改善早期脑损伤过程中大鼠的神经行为学评分,并对血-脑脊液屏障通透性产生影响。Objective To investigate the effects of Neuritin in endoplasmic reticulum stress factor - GRP78 and blood -brain barrier permeability during the process of early brain injury （EBI） after subarachnoid hemorrhage （SAH） of rats. Methods 144 heahhy adult male SD rats were selected from April 2014 to September 2015, and they were divided into control group, Sham group, SAH group and Neuritin group with 36 rats in each group, according to random number table method. Then according to simple random number table method, 6 rats were selected to be executed from each group at time points of 3, 6, 12, 24, 48, 72 h respectively and lateral ventricle injection with Neuritin was given to rates in Neuritin group by stereotactic technique 1 h before execution. 6, 12, 24, 48, 72 h after modeling, Garcia neurological score of rats in each groupwas recorded before their execution; 3, 6, 12, 24, 48, 72 h after modeling, HE staining was used to observe the SAH establishment situation, Meilan was microinjected into lateral ventricles to identify Neuritin injection condition, we detected the protein expression of endoplasmic reticulum stress factor GRP78 in cerebral cortex of rats by Western blotting method, and the Evans blue （EB） content in brain of rats by formamide immersion method to evaluate the blood - brain barrier permeability. Results 6, 12, 24, 48, 72 h after modeling, Garcia neurological scores of rats in SAH group and Neuritin group were lower than those of control group and Sham group （P 〈 0. 05） ; 6, 12, 24, 48, 72 h after modeling, Garcia neurological score of rats in Neuritin group was higher than that of SAH group （ P 〈 0. 05 ） . Hematocele was scattered in the surface of brain of rats in SAH group, and there also was large amount of hematocele in willis ring, cerebellar medulla and ventral brainstem. Meilan circulated around the ventral side of the brain of rats in Neuritin group, while no Meilan in the reverse side of the brain. Intracerebroventrieular injection of rats in Neutritin group was

关 键 词：蛛网膜下腔出血 脑损伤 神经生长因子类 血脑屏障 内质网应激 NEURITIN GRP78 

分 类 号：R743.35[医药卫生—神经病学与精神病学]