梗阻性黄疸大鼠肠黏膜上皮紧密连接蛋白和MLCK的研究  

The study on tight junction proteins and myosin light chain kinase in obstructive jaundice in rats

作　　者：陈振勇[1] 冯贤松[1] 周有生[1] 

机构地区：[1]华中科技大学附属协和医院普通外科,湖北武汉430022

出　　处：《中国普通外科杂志》2008年第2期140-144,共5页China Journal of General Surgery

摘　　要：目的探讨梗阻性黄疸肠黏膜屏障破坏的机制。方法建立梗阻性黄疸大鼠的动物模型,分别于胆管结扎10 d和20 d后,采用免疫组化、Western blot方法检测末端回肠黏膜的紧密连接蛋白成员ZO-1和Occludin与肌球蛋白轻链激酶(MLCK)的分布和表达。结果正常回肠黏膜层ZO-1和Occludin的分布相似,主要位于上皮细胞的边缘,细胞膜顶端,沿绒毛下方均匀连续分布;MLCK主要分布在细胞浆内。梗阻性黄疸时ZO-1和Occludin分布不均,染色变淡,线条模糊,边缘粗糙有毛刺状突起;MLCK分布散乱,染色稀疏。与对照组相比20 d组和10 d组的ZO-1,Occludin,MLCK数量明显减少,强阳性表达率分别从70.0%,80.0%,70.0%降至10 d组的28.6%,28.6%,28.6%(均P<0.05)和20 d组的ZO-1从10 d组的28.6%降至14.3%,35.7%,21.4%(均P<0.05)。20 d组的ZO-1下降较10 d组更为明显(P<0.05),而Occludin和MLCK变化不明显。Western blot检测的结果与之相似。结论梗阻性黄疸时大鼠小肠黏膜上皮ZO-1,Occludin,MLCK分布紊乱,表达下降,小肠黏膜上皮屏障的完整性破坏。Objective To elucidate the mechanisms of disruption of intestinal mucosa barrier in obstructive jaundice. Methods The obstructive jaundice model of rats was set up. At 10 d and 20 d after operation, immunohistochemistry and Western-blot techniques were used to examine the distribution and expression of tight junction proteins （ZO-1, Occludin ） and myosin light chain kinase （MLCK）in intestinal mucosa. Results In normal control groups, the staining of ZO-1 and occludin was predominantly localized to the margins of the epithelial cells and the apex of the cell membrane, and displayed a continuous and uniform distribution along the under surface of the villae. MLCK was distributed mainly in cytoplasm. In obstructive jaundice groups, ZO-1 and occludin staining appeared discontinuous and vague, with rough edges and spiculate processes. The staining of MLCK was also discontinuous and scanty. The strong positive express ratio of ZO- 1 , 0ccludin and MLCK were obviously lower in two experiment group than those in the control group all P 〈0.05）. ZO-1 was significantly decreased in 20 d group than that in 10 d group（ 14.3 % vs28.6 % , P 〈0.05 ）, but no significant difference in occludin and MLCK staining between 10 d group and 20 d group. Similar outcomes were obtained by quantitative analysis of Western blot images. Conclusions Obstructive jaundice rats have distribution of ZO-1 , occludin and MLCK in ileum mucous membrane epithelium, and is associated with breakdown of the integrity of the intestinal barrier.

关 键 词：黄疸 阻塞性/病理学 紧密连接蛋白 肌球蛋白轻链激酶 肠黏膜屏障 

分 类 号：R657.4[医药卫生—外科学]