荧光标记生物素法评价大鼠肠黏膜屏障损伤  

Assessment of intestine barrier injury in rats using fluorescein-labeled biotin

作　　者：张强[1] 李秋荣[1] 王晨阳[1] 顾立立[1] 曲林林[1] 李宁[1] 刘晓祥[2] 黎介寿[1] 

机构地区：[1]南京大学医学院临床学院南京军区南京总医院解放军普通外科研究所,江苏南京210002 [2]南京农业大学作物遗传与种质创新国家重点实验室,江苏南京210095

出　　处：《肠外与肠内营养》2010年第3期153-155,159,共4页Parenteral & Enteral Nutrition

基　　金：国家973项目(2007CB513005);南京军区医学科学技术研究"十一五"计划(06Z40)

摘　　要：目的:利用荧光标记生物素检测大鼠缺血-再灌注肠黏膜屏障损伤。方法:将20只大鼠随机分为对照(假手术)组、缺血-再灌注后1、3和5h组,共四组,每组5只。利用动脉夹夹闭肠系膜上动脉1h后放开,建立大鼠缺血-再灌注模型。观察缺血-再灌注损伤后肠组织形态学改变,并用荧光标记生物素作为探针,应用激光共聚焦显微镜,检测大鼠缺血-再灌注肠黏膜屏障损伤。结果:大鼠缺血-再灌注损伤后,肠黏膜明显受损,病理表现为肠黏膜水肿和炎性细胞浸润。损伤程度随缺血后再灌注时间的延长而增加。激光共聚焦显微镜观察发现,荧光标记生物素穿透缺血-再灌注肠上皮进入黏膜下层,表明肠上皮屏障受到破坏。结论:缺血-再灌注可损伤大鼠肠屏障功能。荧光标记生物素法是一种适合评价实验动物肠黏膜屏障损伤的方法。Objective：To evaluate intestine barrier impairment in ischemia reperfusion （I/R） injury rats using fluorescein-labeled biotin.Methods：Twenty Sprague-Dawley （SD） rats were randomly divided into four groups which were control group （n=5）,and I/R injury groups for 1h,3h and 5h （five animals for each group）.I/R injury was performed by occluding the superior mesenteric artery （SMA） with an atraumatic vascular artery clamp for 1h.The histological alterations were examined by hematoxylin and eosin （H-E） staining,and intestine barrier disruption was investigated by fluorescein-labeled biotin with confocal immunofluorescence microscope.Results：The intestinal mucosa was injuried obviously,including hemorrhage,oedema and inflammatory cell infiltration.And the degree of histological destruction increased as the reperfusion period was prolonged.Moreover,using immunofluorescence microscopy we found that fluorescein-labeled biotin penetrated into the lamina propria,which indicated disruption of intestinal epithelium.Conclusion：The intestine barrier function is impaired after I/R injury.And fluorescein-labeled biotin is a suitable method to assess intestine barrier lesion in I/R injury rats.

关 键 词：缺血-再灌注 荧光标记生物素 肠黏膜屏障损伤 

分 类 号：R363[医药卫生—病理学]