摘要
目的通过分别建立SD大鼠完全脑缺血和脑缺血再灌注模型,比较研究完全脑缺血和脑缺血再灌注介导的神经元损伤,以及相关炎症因子的表达。方法将成年雄性SD大鼠(42只,体重250～350 g)随机分为7组,构建脑缺血24、48、72 h模型及脑缺血2 h再灌注24、48、72 h模型。以Longa评分标准对大鼠进行神经功能评分;2,3,5-氯化三苯基四氮唑(2,3,5-triphenyltetrazolium chloride,TTC)染色测定脑梗死体积;苏木精-伊红(hematoxylin-eosin,HE)染色检测大脑皮层、海马CA3区神经元损伤程度;免疫荧光法检测NOD样受体蛋白3(NOD-like receptor protein 3,NLRP3,又称NALP3)在脑组织中的表达及定位。结果与正常组比较,完全脑缺血和脑缺血再灌注模型大鼠的Longa评分更高,且脑组织梗死体积显著增大。完全脑缺血和脑缺血再灌注模型大鼠大脑皮层和海马CA3区的神经元损伤明显;随着再灌注时间的延长,再灌注48 h大鼠的脑皮层和海马CA3区神经元损伤明显减轻,但在再灌注72 h时再度加重。与正常组比较,完全缺血24 h和缺血2 h再灌注24 h后,NALP3-Cy3荧光表达明显增强。结论完全脑缺血和脑缺血再灌注均可造成大脑皮层和海马CA3区的神经元损伤,仅在缺血2 h再灌注48 h时大脑损伤可部分恢复;在上述两模型中炎症因子NALP3的表达均显著升高。
Objective Models of complete cerebral ischemia and cerebral ischemia reperfusion in SD rats were established to compare and study the neuronal cell damage and the expression of related inflammatory factors. Methods Adult male SD rats(80 rats,weighing 250-350 g)were randomly divided into seven groups.Longa scores were used to assess neurological function in rats.TTC staining was used to measure the volume of cerebral infarction.HE staining was used to detect the degree of neuronal damage in cerebral cortex and hippocampal CA3 region.Immunofluorescence was used to detect the expression level and expression site of NALP3 protein. Results The Longa score was higher in the models of complete cerebral ischemia and cerebral ischemia reperfusion than in the normal group.Meanwhile,the infarct size of the brain tissue was significantly increased.The results of HE staining showed that rats with complete cerebral ischemia and rats with cerebral ischemia-reperfusion had significant neuronal damage in the cerebral cortex and hippocampal CA3 region.Moreover,with the extension of reperfusion time,neuronal damage in the cortex and hippocampal CA3 area of rats was significantly reduced 48 h after reperfusion,but re-emerged 72 h after reperfusion.Immunofluorescence analysis indicated that the expression of NALP3-Cy3 was significantly increased in the 24 h complete ischemia group and 2 h ischemia/24 h reperfusion group,compared with the normal group. Conclusion Complete cerebral ischemia and cerebral ischemia reperfusion can mediate neuronal damage in the cerebral cortex and hippocampal CA3 region,and brain damage can be partially restored only in 2 h ischemia/48 h reperfusion group.The expression of the inflammatory factor NALP3 is significantly increased in both constructed models.
作者
申鑫
张志兵
陈雅梦
王恒琦
张宏伟
柳赟昊
张玲
曹晓璐
Shen Xin;Zhang Zhibing;Chen Yameng(Key Laboratory of Occupational Hazard Identification and Control in Hubei Provincial,Department of Environmental Health and Occupational Medicine,School of Public Health,Wuhan University of Science and Technology,Wuhan 430065,China)
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2019年第2期200-205,216,共7页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.81801307
81300536
81671514)
