细胞凋亡在急性坏死型胰腺炎早期肠黏膜上皮细胞死亡中的作用  

Role of apoptosis in the cell death of rat intestinal epithelium during the early stage of acute necrotizing pancreatitis

作　　者：王兴鹏[1] 王冰娴[1] 吴恺[1] 徐选福[1] 

机构地区：[1]上海市第一人民医院消化科,消化病研究室,200080

出　　处：《中华消化杂志》2001年第5期267-270,共4页Chinese Journal of Digestion

基　　金：上海市科技启明星培养计划资助! (99QB14 0 0 9)

摘　　要：目的　观察急性坏死型胰腺炎 (ANP)大鼠早期肠黏膜上皮细胞凋亡的发生及其演变规律 ,探讨其在肠黏膜屏障功能障碍中的作用。方法　Spraque Dawley大鼠 48只 ,采用胆胰管内逆行注入5 %牛磺胆酸钠溶液诱导大鼠ANP模型。假手术组 (shamoperation ,SO)大鼠仅作剖腹术。术后 3、6、12和 2 4h分批处死大鼠 ,取末端回肠组织 ,分别应用DNA琼脂糖凝胶电泳 ,异硫氰基荧光素 (FITC)结合的Annexin V和碘化丙啶 (PI)标记细胞作流式细胞仪 (FCM)检测和免疫组化 (TUNEL法 )等技术 ,研究ANP大鼠肠黏膜上皮细胞凋亡。结果　DNA琼脂糖凝胶电泳结果显示 ,SO组仅于 12h出现“梯形(ladder)”条带 ;ANP大鼠 ,各时点均可见“梯形”条带。FCM检测结果表明 ,术后 3、6、12和 2 4hSO组肠黏膜上皮脱落细胞凋亡比例分别为 (5 3 .7± 3 .7) %、(2 7.6± 6 .0 ) %、(39.0± 4.8) %和 (2 9.0± 11.3) % ;ANP组分别为 (5 0 .3± 11.3) %、(79.7± 9.2 ) %、(4 7.8± 17.3) %和 (4 9.6± 9.5 ) % ,术后 6h达峰值 ,且较SO组明显增高 (P <0 .0 1) ,凋亡与坏死细胞 (A/N)比值为 2 7.7± 17.9。TUNEL法显示 ,凋亡细胞呈细胞核固缩、断片状 ,术后 3、6、12和 2 4hSO组凋亡指数为 6 .2± 2 .4、7.5± 1.7、10 .5± 0 .9和 5 .3± 0 .8;ANP组为Objective To investigate whether apoptosis of intestinal epithelial cells occurs at the early stage of acute necrotizing pancreatitis (ANP) in rats. Methods Fourty eight Spraque Dawley rats were used. ANP model of rats was induced by retro injection of 5% sodium taurocholate into biliopancreatic duct. Laparotomized animals without induction of ANP (sham operation) served as controls. The distal segment of ileum specimens were harvested at 3 h, 6 h, 12 h and 24 h after operation and the apoptosis of intestinal epithelial cells was studied by DNA gel electrophoresis, FITC conjugated annexin V and propidium iodide (PI) staining cells analyzed by Flow Cytometry (FCM) and immunohistochemical procedures (TUNEL method). Results The DNA electrophoresis showed that typical “ladder” patterns appeared at all indicated time points in ANP group, while the DNA specimens from control group presented a single chromosomal lane, except the one at 12 h. Apoptotic percentage of detached intestinal epithelial cells assayed using Annexin V kit by FCM were (53.7±3.7)%, (27.6±6.0)%, (39.0±4.8)%, (29.0±11.3)% at 3 h, 6 h, 12 h and 24 h in control group and (50.3±11.3)%, (79.7±9.2)%, (47.8±17.3)%, (49.6±9.5)% in ANP group. There was a significant difference between two groups at 6 h, P <0.05. The ratio of apoptotic cells and necrotic cells (A/N) was 27.7±17.9. Apoptotic index determined by TUNEL method were significantly increased in ANP group (17.5±3.5, 20.4 ±2.9, 14.8±1.7, 14.2±2.1) compared with control group (6.2±2.4, 7.5±1.7, 10.5±0.9, 5.3 ± 0.8 ) during all observed period, P <0.01, peaked at 6 h. Conclusions It was suggested that apoptosis of intestinal epithelial cells increased at early stage of ANP in rat, especially at 6 h. Apoptosis is a principle model of cell death in rat intestinal epithelium of ANP. These might be involved in the development of intestinal barrier failure during the course of ANP.

关 键 词：急性胰腺炎 肠黏膜屏障 细胞凋亡 上皮细胞死亡 

分 类 号：R657.51[医药卫生—外科学]