胆汁外引流对梗阻性黄疸大鼠肠黏膜紧密连接影响的实验研究  

Effect of External Biliary Drainages on Tight Junctions of Intestinal Epithelia in Rats with Obstructive Jaundice

作　　者：陈振勇[1] 冯贤松[1] 杨鹏[1] 周有生[1] 

机构地区：[1]华中科技大学附属协和医院综合外科,武汉430022

出　　处：《中国普外基础与临床杂志》2010年第5期454-458,共5页Chinese Journal of Bases and Clinics In General Surgery

摘　　要：目的探讨胆汁在保护小肠黏膜屏障中的作用及其机理。方法 50只Wistar大鼠随机分为对照组(n=10)、梗阻性黄疸(阻黄)组(n=20)及外引流组(n=20)。造模10 d后测定血浆内毒素水平;取末端回肠黏膜组织,光镜下观察黏膜形态改变并测量相关指标,采用免疫组化及Western blot法检测紧密连接相关蛋白(闭锁小带蛋白-1及闭锁蛋白)的表达。结果阻黄组小肠黏膜萎缩明显,其肠绒毛高度、黏膜厚度和隐窝深度与对照组相比分别下降27.8%、21.7%和25.4%(P=0.001、0.001、0.040);外引流组各指标较对照组无明显变化(P=0.050、0.070、0.080),而明显优于阻黄组(均P=0.001)。阻黄组的血浆内毒素水平高达(1.49±0.27)EU/ml,明显高于对照组的(0.27±0.09)EU/ml(P=0.001);外引流组为(0.91±0.25)EU/ml,高于对照组而低于阻黄组(P=0.001)。免疫组化结果显示,闭锁小带蛋白-1的强阳性表达从对照组的7/10例降至阻黄组的6/20例(P=0.040),闭锁蛋白则从8/10例降至7/20例(P=0.020);而外引流组分别为8/20例(P=0.100、0.210)和9/20例(P=0.060、0.200),与前2组相比差异均没有统计学意义。Western blot分析显示,阻黄组闭锁小带蛋白-1和闭锁蛋白的表达水平较之对照组均明显下降(P=0.001、0.010);外引流组高于阻黄组(P=0.005、0.014),而闭锁小带蛋白-1的表达水平低于对照组(P=0.001),闭锁蛋白的表达与对照组比较差异无统计学意义(P=0.062)。结论肠道内胆汁缺乏会破坏肠紧密连接相关蛋白的成分,损伤肠黏膜屏障;胆道梗阻时由于存在其他因素,肠黏膜屏障损伤更严重。胆汁在保护肠黏膜屏障中具有重要作用。Objective To investigate the effects and mechanisms of bile on small intestine mucosal barrier.Methods Fifty Wistar rats were assinged into 3 groups randomly： obstructive jaundice（OJ） group（n=20）,biliary external drainage group（n=20） and control group（n=10）.Ten days after operation,the plasma endotoxin level was determinated,the terminal ileum mucosas was obtained to be morphologically measured by light microscope,and immunohistochemistry and Western blot were uesd to examine the expressions of tight junction proteins zona occludens-1（ZO-1） and occludin in the mucosas.Results Atrophy significantly appeared in the distal ileum mucosas in OJ group.Compared with control group,the intestinal villus height,mucosa thickness and crypt depth in OJ group were obviously decreased 27.8%,21.7%,and 25.4%（P=0.001,0.001,0.040）.There were no differences between external drainage group and control group（P=0.050,0.070,0.080）;While the values of external drainage group were significantly higher than those in OJ group（all P=0.001）.The level of plasma endotoxin was up to（1.49±0.27） EU/ml in OJ group compared with control group（（0.27±0.09） EU/ml）,P=0.001.In external drainage group,the value was（0.91±0.25） EU/ml,which was obviously higher than that in control group and lower than that in OJ group（all P=0.001）.Immunohistochemical study showed strong positive expression of ZO-1 dropped from 7/10 in the control group to 6/20 in OJ group（P=0.040）,occludin expression was 8/10 in control group and 7/20 in OJ Group（P=0.020）;expressions of them in external drainage group（8/20（P=0.100,0.210） and 9/20（P=0.060,0.200）） displayed no significant differences compared with the other twogroups.Quantitative testing of Western blot showed the expressions of ZO-1 and occludin in OJ group were significantly lower than those in control group（P=0.001,0.010）,the values in external drainage group were higher than those in OJ group（P=0.005,0.014）.The expression of ZO-1 was lowe

关 键 词：梗阻性黄疸 胆汁 肠黏膜屏障 闭锁小带蛋白-1 闭锁蛋白 

分 类 号：R657.43[医药卫生—外科学]