灭活血吸虫卵对实验性结肠炎肠黏膜的影响  

Effect of inactivated schistosome ova on intestinal epithelial tight junctions during tinitrobenzene sulfonic acid-induced colitis

作　　者：夏晨梅 张顺财[2] 

机构地区：[1]温岭市第一人民医院消化科,浙江省317500 [2]复旦大学附属中山医院消化科

出　　处：《中华消化杂志》2010年第2期81-86,共6页Chinese Journal of Digestion

摘　　要：目的研究灭活血吸虫卵对2，4，6-三硝基苯磺酸（tinitr。benzenesulfonicacid，TNBS）诱导小鼠结肠炎肠黏膜紧密连接蛋白z0—1和Oceludin基因及蛋白表达影响及其机制。方法清洁级BAI。B／C雌性小鼠50只分成对照组（10只）、TNBS＋0．9％氯化钠溶液组（20只）和TNBS＋血吸虫卵组（20只）。TNBS＋血吸虫卵组在造模前第3、14天分别给予腹腔注射冰冻灭活血吸虫卵10000个（1ml冰0．9％氯化钠溶液混悬液）；TNBS＋0．9％氯化钠溶液组给予相同体积的冰0．9％氯化钠溶液腹腔注射。后两组予TNBS溶液灌肠（100mg／kg）建立结肠炎模型，建模后第7天处死存活小鼠，观察各组小鼠结肠的大体形态和HE染色光镜下病理特征；荧光实时定量PCR法测定结肠组织的Occludin和ZO-1基因表达；Western印迹法检测蛋白表达；免疫组化法测定结肠组织紧密连接蛋白表达分布。结果TNBs＋血吸虫卵组小鼠死亡率较TNBS＋0．9％氯化钠溶液组明显下降（15％比30％）。TNBS＋0．9％氯化钠溶液组组织学评分为（4．21±0．40）分，较TNBS＋血吸虫卵组和对照组高[（1．74±0．10）和（1．06±0．20）分，P〈0．05]。TNBS＋0．9％氯化钠溶液组ZO-1和0ccludinmRNA表达量较对照组显著下降（P〈0．01），而TNBS＋血吸虫卯组较TNBS＋0．9％氯化钠溶液组显著增加（P〈0．05）。TNBS＋0．9％氯化钠溶液组ZO-1蛋白相对灰度值较正常对照组降低50．3％（P〈0．05），而TNBS＋血吸虫卵组较TNBS＋0．9％氯化钠溶液增加41．1％（P〈0．05）；TNBS＋0．9％氯化钠溶液组Occludin相对灰度值较对照组下降48．7％（P〈0．05），而血吸虫卵组较TNBS＋0．9％氯化钠溶液组增加23．6％（P〈0．05）。ZO-1、Occludin蛋白染色强度TNBS＋0．9％氯化钠溶液组分布均较对照组间增强（P〈0．01），而TNBS＋血吸虫卵组染色强度信号分布较TNBS＋0．9％氯化钠溶液组�Objective To observe the possible effect of inactivated sehistosome ova on the expression of intestinal epithelial tight junctions ZO-1 and Occludin gene in mouse colitis induced by tinitrobenzene sulfonie acid （TNBS） and its mechanism. Methods Fifty mice were divided into control group （group A, n= 10）, TNBS plus normal saline（NS） group（group B, n = 20） and TNBS plus inactivated schistosome ova group（group C, n= 20）. Group C was exposed to 10 000 freeze-killed schistosome ova by intraperitoneal injection at day 14 and day 3 before colitis induction. Meanwhile, group B was exposed to I ml NS by intraperitoneal injection. The mice in group B and C were challenged with 3 mg TNBS to induce colitis. All mice were killed 7-day after colitis induction and assessed with following variables including mortality, pathological change with HE staining of colon.The transcription levels of ZO-1 and Occludin in colon tissues were examined using Real-time PCR. The expression and distribution of ZO-1 and Occludin proteins were detected by Western blotting and immunohistoehemistry. Results In comparison with group B inactivated schistosome ova most effectively reduced the mortality （30 % vs 15 %） and histopathologic severity of TNBS-induced colitis （4. 21 ± 0. 40 vs 1. 74 ±0. 10）. The transcription levels of ZO-1 and Occludin in group B were decreased compared with those in group A and group C （P〈0.01）. When compared with group B, group C showed a significant elevation of the alteration of ZO-1, Occludin proteins expression and localization. Conclusion The results clearly show that schistosome ova treatment reduced the severity of experimental colitis through the regulation of tight junction proteins.

关 键 词：炎症性肠病 血吸虫属 紧密连接蛋白 模型 动物 

分 类 号：R574.6[医药卫生—消化系统]