肠缺血/再灌注损伤对大鼠肠胶质细胞影响的实验研究  

Effects of intestinal ischemia / reperfusion injury on enteric glia cells in rats

作　　者：杜明华[1] 罗红敏[2] 石现[1] 喻文[2] 马丽[2] 林志龙[2] 王欢[1] 胡森[2] 

机构地区：[1]解放军总医院,北京100853 [2]解放军总医院第一附属医院烧伤研究所休克与多器官障碍实验室,北京100048

出　　处：《感染．炎症．修复》2012年第4期203-206,F0002,共5页Infection Inflammation Repair

基　　金：国家973项目(2012CB518101)

摘　　要：目的:探讨大鼠肠缺血/再灌注损伤后肠胶质细胞的变化情况,为研究其在消化道疾病中的作用提供实验基础。方法:采用夹闭肠系膜上动脉60 min然后再恢复血流60 min或120 mim的方法建立肠缺血/再灌注损伤模型,肠组织切片行HE染色,显微镜下观察;应用改进的方法制作肠肌问神经丛全层铺片,S100β免疫组化和荧光染色法显示肠肌间神经丛胶质细胞,荧光显微镜下观察、拍照,应用Image-pro Plus 5.0图像分析系统,计算S100β阳性细胞数量。结果:缺血60 min/再灌注60 min或120 min,肠HE染色可见绒毛变钝、大量炎性细胞浸润;采用改进的方法制作肠肌间神经丛全层铺片并行S100β免疫组化和荧光染色,可以清楚地显示肌间神经丛肠胶质细胞结构;缺血/再灌注后肠胶质细胞明显减少,其中缺血60 min/再灌注120 min更显著(P<0.05)。结论:肠道肌间神经丛铺片结合免疫组化方法是研究肠道肌间神经丛胶质细胞的理想方法,肠肌间神经丛胶质细胞在缺血/再灌注过程中明显减少,缺血/再灌注过程中肠黏膜的损伤可能与肠胶质细胞有关。Objective：To investigate the changes in enteric glia cells （EGCs）in the rat model of intestinal ische-mia/reperfusion injury. Methods：Aduh male rats were subjected to intestinal ischemia for 60 minutes by occluding the superior mesenteric artery, and followed by reperfusion. Small intestinal segments were resected at 60 minutes or 120 minutes after reperfusion and observed after HE staining with optic microscope. The whole mount prepara-tions of myentric plexus were obtained by the improved method, and EGCs of myentric plexus were observed by immunohistochemistry and immunofluoreseence techniques. Image-pro Plus 5.0 system were used to count S100β positive cells （EGCs）. Results：Pathological changes in intestinal tissue showed damage of mucosal epithelial cells, shown as bluntness of intestinal villi and infiltration of inflammatory ceils after 60-minute ischemia followed by 60 or 120-minute reperfusion. The structure of EGCs of myenteric plexus was clear with the improved method for whole mount preparations of myentric plexus, immunohistochemistry, and immunofluorescence technique of S100β staining. The results showed that EGCs decreased obviously after ischemia/reperfusion, especially after 120 mi-nutes reperfusion （P〈0.05）. Conclusion： The improved whole mount preparations, combined with immunohisto-chemistry techniques,are really good methods to show EGCs in myenteric plexus. Ischemia/reperfusion injury might lead to loss of EGCs, which is related to the intestinal mucosa injury.

关 键 词：小肠 缺血 再灌注损伤 肠胶质细胞 肠肌间神经丛铺片 免疫组化 免疫荧光 

分 类 号：R541[医药卫生—心血管疾病]