外周血白细胞凋亡障碍与大鼠肠缺血-再灌注损伤的关系  

Correlation of peripheral leukocyte apoptosis insufficiency and intestinal injury following mesenteric ischemia/reperfusion in rats

作　　者：莫燕娜[1] 王丹妹[1] 吉丽敏[1] 翁阳[2] 何佟[1] 陈世民[1] 

机构地区：[1]海南医学院机能学实验室,海南海口571101 [2]海南医学院病理学教研室,海南海口571101

出　　处：《中国病理生理杂志》2009年第2期356-360,共5页Chinese Journal of Pathophysiology

基　　金：海南省自然科学基金资助项目(No.30625);海南省普通高校优秀中青年教师科研与教学奖励基金资助项目

摘　　要：目的:探讨外周血白细胞和中性粒细胞凋亡障碍在肠缺血-再灌注(IR)损伤中的作用。方法:20只雄性SD大鼠随机分为肠IR组和假手术对照组,每组10只。肠IR组夹闭肠系膜上动脉(SMA)30min后再灌注60min;对照组只分离而不夹闭SMA。观察肠黏膜形态学变化;检测肠黏膜上皮细胞凋亡指数(TUNEL法)和caspase-3活性;Annexin-V/PI法检测外周血白细胞和中性粒细胞(PMN)凋亡比例;检测夹闭前、夹闭30min、再灌注30min、再灌注60min外周血白细胞数量。结果:(1)光镜下,对照组未见肠黏膜损伤,而IR组肠黏膜损伤严重;(2)IR组肠黏膜上皮细胞凋亡指数和caspase-3活性比对照组明显增高(P<0.05);(3)IR组外周血白细胞和PMN凋亡比例比对照组明显降低(P<0.05);IR组外周血白细胞数量在缺血30min时明显增高,再灌注后进一步增高,与缺血前和对照组比都有显著差异(P<0.05)。(4)肠黏膜caspase-3活性与外周血白细胞凋亡比例有显著的负相关(r=-0.764,P<0.01),与PMN凋亡比例也有显著的负相关(r=-0.845,P<0.01);肠黏膜上皮细胞凋亡指数与PMN凋亡比例也有显著的负相关(r=-0.638,P<0.05)。结论:外周血白细胞数量增加及凋亡障碍与缺血-再灌注引起的肠黏膜细胞损伤密切相关。AIM： To investigate the role of peripheral blood leukocytes and polymorphonuclear neutrophils （PMNs） on intestinal injury following mesenteric ischemia/reperfusion （IR） in rats. METHODS： Twenty adult, male Sprague - Dawley rats, weighing 200 - 230 g, were randomly divided into two groups. The control group （CON） consisting of 10 rats was subjected to laparotomy and separation of superior mesenteric artery （SMA） only. The ischemia/reperfusion （IR） group consisting of 10 rats, was subjected to laparotomy, followed by occlusion of the superior mesenteric artery （SMA） by an atraumatic microvascular clamp for 30 min. At the end of ischemic period in IR, the microvascular clamp was removed and the intestinal segment was reperfused for 60 min. The pathological changes of the ileal mucosal tissue were evaluated. The apoptosis of intestinal mucosal epithelial cells was examined by terminal deoxylnucleotidy - 1 transferase mediated- dUTP nick end- labeling （TUNEL）. The enzymatic activity of casapse- 3 in mucosal cells was determined using a colorimetric assay. The percentages of apoptotic peripheral blood leukocytes and PMNs were measured by flow cytometry using Annexin - V/PI double staining assay. The numbers of peripheral blood leukocytes in each animal was measured at baseline, 30 min of ischemia, and 30 min and 60 min of reperfusion. RESULTS ： （ 1 ） Compared to CON group animals,the most severe mucosal injury was observed in IR group under optical microscope. （2） The number of apoptotic mucosal epithelia cells and enzymatic activity of caspase - 3 were significantly higher in IR than those in CON group （ P 〈 0. 05 ）. （3） The percentages of apoptotic peripheral blood leukocytes and PMNs were significantly lower in IR group than those in CON group （ P 〈 0. 05 ）. The number of peripheral blood leukocytes in IR group was increased obviously after SMA closed for 30 min （ P 〈 0. 05 ）, and was higher following reperfusion （ P 〈 0. 05 ）, and significantly h

关 键 词：肠 再灌注损伤 白细胞凋亡 半胱氨酸天冬氨酸蛋白酶3 

分 类 号：R363[医药卫生—病理学]