Necrostatin-1对大鼠脑缺血再灌注损伤后小胶质细胞和炎症因子的影响  

Influence of necrostatin-1 in microglia activation and inflammatory factor levels after cerebral ischemia/reperfusion damage in rats

作　　者：支中文 杨龙[1] 杜波[1] 周延龙[1] 耿德勤[1] 程言博[1] 

机构地区：[1]徐州医学院附属医院神经内科,徐州221002

出　　处：《中华神经医学杂志》2015年第8期757-763,共7页Chinese Journal of Neuromedicine

摘　　要：目的探讨干预程序性坏死途径对大鼠脑缺血再灌注损伤后小胶质细胞活化及相关炎症因子的影响及意义。方法采用改良线栓法构建大鼠右侧大脑中动脉缺血再灌注模型。（1）SD大鼠按随机数字表法分为脑缺血再灌注6h组、脑缺血再灌注12h组、脑缺血再灌注24h组、脑缺血再灌注72h组，每组6只。免疫组化染色检测不同时间点小胶质细胞激活标志离子型钙接头蛋白-1（iba-1）的表达，根据实验结果选取iba-1表达相对明显的时间点。（2）SD大鼠按随机数字表法分为假手术组、脑缺血再灌注组、Necrostatin-1（Nec-1）（80nm01）干预组、Nec-1（160nmol）干预组．每组6只，并于脑缺血后2h给予Nec-1干预。采用Longa法进行神经功能损伤评分、TTC法检测梗死体积，根据实验结果选取Nec-1干预的适宜给药剂量。（3）SD大鼠按随机数字表法分为假手术组、脑缺血再灌注组、溶剂组、Nec-1干预组，每组6只，并于脑缺血后2h给予Nec．1或DMS0溶剂干预。HE染色观察梗死周围组织神经元存活情况，免疫组化染色检测梗死周围组织iba-l的表达．免疫组化染色及Westernblotting分别检测炎症因子肿瘤坏死因子-α（TNF-α）、白介素-1β（IL-1β）、胶质源性神经营养因子（GDNF）免疫阳性细胞及蛋白的表达。结果（1）iba-1表达在脑缺血再灌注24h后升高明显，与其他各时间点比较差异有统计学意义（（P〈0．05）。（2）与脑缺血再灌注组相比，两Nec．1干预组神经功能损伤评分明显降低、梗死体积明显减小，差异均有统计学意义（P〈0．05）；与Nec-1（80nmoll干预组相比，Nec-1（160nmol）干预组神经功能损伤及梗死体积的改善效果更为明显，差异均有统计学意义（氏0．05）。（3）HE染色显示脑缺血再灌注组梗死周围组织炎症细胞浸润、神经元减少、胞体皱缩、细胞核固缩；与脑缺血再灌注组相比，Objective To investigate the influence of necroptosis pathways in microglia activation and inflammatory factor levels in cerebral ischemia/reperfusion （I/R） injury in rats and their significances. Methods A modified suture method was used to establish the models of middle cerebral artery FR in rats. （1） SD rats according to the random number table were divided into cerebral UR 6 h group, cerebral I/R 12 h group, cerebral FR 24 h group, and cerebral I/R 72 h group （n=6）; the expression of ionized calcium binding adaptor molecule-1 （iba-1） was detected by immunohistochemical staining, and time points enjoyed obvious iba-1 expression were selected according to the experimental results. （2） SD rats were randomly divided into sham operation group, cerebral I/R group, 80 nmolnecrostatin-1 （Nec-1） intervention group and 160 nmol Nec-1 intervention group （n=6）, and the Nec-1 intervention was given 2 h after ischemia; Longa method was used to evaluate the neurological function scores and TTC method was used to detect the infarct volume; and the appropriate dosages of Nec-1 were selected according to these results. （3） SD rats were randomly divided into sham operation group, cerebral I/R group, solvent group and Nec-1 intervention group （n=6）, and Nec-1 or DMSO intervention was given 2 h after ischemia. HE staining was used to observe the survival and proliferation of microglias around the infarction tissues; immunohistochemical staining was used to detect the iba-1 expression surrounding the infarction tissues; immunohistochemistry and Western blotting were employed to observe the cytokine tumor necrosis factor （TNF）-α and interleukin （IL）-1β and glia-derived neurotrophic factor （GDNF） expressions. Results （1） The iba-1 expression at cerebral I/R 24 h group was significantly increased as compared with that in other groups （P〈0.05）. （2） As compared with those in the cerebral I/R group, the neurological function scores and infarct volume were significantly decr

关 键 词：程序性坏死 Necrostatin-1 脑缺血再灌注损伤 小胶质细胞 肿瘤坏死因子-α 白介素-1Β 胶质源性神经营养因子 

分 类 号：R743.3[医药卫生—神经病学与精神病学]