急性脑缺血/再灌注后细胞病理改变及星形胶质细胞活化规律  

Research on cellular damages and astrocyte activation after cerebral ischemia and reperfusion

作　　者：兰瑞[1] 马云枝[1] 朱世瑞[1] 付雪琴 邹旭欢 王漫漫 王玮玮 王保奇[1] 张勇[3] LAN Rui;MA Yun-zhi;ZHU Shi-rui;FU Xue-qin;ZOU Xu-huan;WANG Man-man;WANG Wei-wei;WANG Bao-qi;ZHANG Yong(the First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China;the First Clinical Medical School of Henan University of Chinese Medicine,Zhengzhou 450000,China;the Third Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)

机构地区：[1]河南中医药大学第一附属医院,河南郑州450000 [2]河南中医药大学第一临床医学院,河南郑州450000 [3]郑州大学第三附属医院,河南郑州450000

出　　处：《中国药理学通报》2023年第4期739-744,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81973618,81503422);河南省自然科学基金资助项目(No 202300410399)。

摘　　要：目的观察脑缺血/再灌注后不同时间点细胞病理改变及星形胶质细胞活化的规律。方法本实验共采用85只SD雄性SPF级大鼠,将其随机分为假手术组、大脑中动脉闭塞90 min再灌注不同时间点组,后者分为再灌注3、6、12、24、48 h组。HE染色观察细胞病理改变,采用透射电镜观察脑缺血/再灌注后星形胶质细胞的亚显微结构,免疫印迹联免疫荧光法检测脑缺血/再灌注损伤后的GFAP表达。结果脑缺血/再灌注后不同时间点可导致神经细胞发生不同程度的缺血/再灌注损伤,再灌注后24 h细胞损伤最为严重。透射电镜结果提示星形胶质细胞亚显微结构发生改变,细胞出现不同程度肿胀,线粒体嵴断裂等。脑缺血/再灌注后在缺血区GFAP表达呈时间依赖性逐渐升高,再灌注48 h后表达最高,而在梗死区表达逐渐减少。结论脑缺血/再灌注后24 h神经细胞损伤最重,缺血半暗带区的星形胶质细胞形态发生变化,星形胶质细胞激活,伴随再灌注的推移,梗死区、缺血区边界逐渐清晰,可能出现胶质瘢痕。Aim To observe cellular damage and astrocyte activation at different time points of cerebral ischemia and reperfusion.Methods The middle cerebral artery of male SpragueDawley rats was occluded for 90 min followed by different time points of reperfusion.Eighty-five SPF male SD rats were randomly divided into control group(Sham),IR3,6,12,24 and IR48h(MCAO followed by 48 h of reperfusion)group.Cerebral ischemia and reperfusion injury was observed by HE staining,and the structure of astrocytes was estimated with transmission electron microscopy(TEM).GFAP expression was detected by immunofluorescence staining and Western blot.Results Cerebral ischemia following by different time points of reperfusion led to different degrees of cellular damage,which was the most serious at 24 h of reperfusion.TEM showed destruction of astrocytes structure,swollen organelles and broken mitochondrial ridge.After cerebral ischemia-reperfusion,the expression levels of GFAP were significant up-regulated in the ischemic penumbra cortex and the highest was at 48 h of reperfusion,indicating astrocytes were activated.In addition,the results showed the gradual decrease in GFAP expression in the infarct core.Conclusions After cerebral ischemia-reperfusion,cellular damage is aggravated,and astrocytes are gradually activated in the ischemic penumbra.With the extension of reperfusion time,the boundaries of infarct area and ischemic area are gradually clear,and scarring may occur.

关 键 词：脑缺血/再灌注 急性期 星形胶质细胞活化 细胞损伤 GFAP 胶质瘢痕 

分 类 号：R-332[医药卫生] R322.81R329.24R743.31