梗阻性黄疸时脾切除对肠黏膜屏障影响的实验研究

目的 探讨脾脏在梗阻性黄疸(阻黄)中对肠黏膜屏障的作用及其机制.方法 50只Wistar大鼠随机分组,阻黄组开腹结扎胆总管;阻黄+脾切除组,同时切除脾脏.术后7d观察血浆内毒素水平的变化,用乳果糖/甘露醇(L/M)比值检测肠黏膜通透性;采用免疫组织化学、Western印迹检测末端回肠紧密连接蛋白闭锁小带-1(ZO-1)、闭锁蛋白的表达,并利用图像分析系统对Western印迹图像进行定量分析.结果 阻黄+脾切除后L/M的比值和血浆内毒素水平较阻黄组明显下降(均P=0.001).与阻黄组相比,阻黄+脾切除组的平均肠绒毛高度和隐窝深度有所上升(P=0.019、0.001).免疫组化显示术后7 d阻黄组ZO-1蛋白强阳性表达数(6/18)下降明显(P=0.021),阻黄+脾切除组(8/17)染色较阻黄组变化不大;闭锁蛋白的染色阻黄+脾切除组强阳性表达(7/17)高于阻黄组(4/18)(P=0.026).通过对Western印迹图像进行定量分析也得出同样的结论.结论 阻黄后肠黏膜通透性增加,肠黏膜屏障受损.同时切除脾脏,肠紧密连接蛋白成分的数量和分布改变,肠黏膜屏障的损害减轻.

Abstract：

Objective To investigate the effects of splenectomy on the intestine mucosa barrier in rats with obstructive jaundice. Methods 50 Wistar rats were divided randomly into the obstructive jaundice group (OJ), in which the animals underwent operation to ligate common bile duct, and the obstructive jaundice + splenectomy group (OJ+ S). Seven days post-operation, plasma endotoxin levels were detected. Intestinal mucosa permeability was measured by the ratios of lactulose and mannitol (L/M). Immunohistochemistry and Western blot were used to examine the expression of tight junction proteins (ZO-1 and occludin) in the distal ileum mucosa. Western blots images were analyzed quantitatively. Results Average ratios of L/M and plasma endotoxin were decreased obviously in the OJ+S group compared to those in the OJ group (all P=0. 001). Compared with the OJ group, the average intestinal villus height and mucosa thickness were upgraded somewhat in the OJ + S group (P = 0.019, 0. 001 ). By immunohistochemistry staining seven days post-operation, same comment as above the amounts of strong positive expression of ZO-1 were significantly decreased in the OJ group (6/18, P-0. 021). There wewas no difference between the OJ+S group(8/17) and the OJ group.The amount of strong positive expression of occludin was higher in the OJ + S group than that of the OJ group(10/17 vs 4/18, P= 0. 026). The same outcomes were obtained by quantitative Western blot images. Conclusion The intestinal epithelial permeability was increased in rats with obstructive jaundice,and intestinal barrier was damaged. After excising spleen, the amount and distribution of tight junction proteins were changed and the impairment of intestinal barrier was abated.

The effects of splenectomy on intestinal mucosa barrier in rats with obstructive jaundice

Objective To investigate the effects of splenectomy on the intestine mucosa barrier in rats with obstructive jaundice. Methods 50 Wistar rats were divided randomly into the obstructive jaundice group (OJ), in which the animals underwent operation to ligate common bile duct, and the obstructive jaundice + splenectomy group (OJ+ S). Seven days post-operation, plasma endotoxin levels were detected. Intestinal mucosa permeability was measured by the ratios of lactulose and mannitol (L/M). Immunohistochemistry and Western blot were used to examine the expression of tight junction proteins (ZO-1 and occludin) in the distal ileum mucosa. Western blots images were analyzed quantitatively. Results Average ratios of L/M and plasma endotoxin were decreased obviously in the OJ+S group compared to those in the OJ group (all P=0. 001). Compared with the OJ group, the average intestinal villus height and mucosa thickness were upgraded somewhat in the OJ + S group (P = 0.019, 0. 001 ). By immunohistochemistry staining seven days post-operation, same comment as above the amounts of strong positive expression of ZO-1 were significantly decreased in the OJ group (6/18, P-0. 021). There wewas no difference between the OJ+S group(8/17) and the OJ group.The amount of strong positive expression of occludin was higher in the OJ + S group than that of the OJ group(10/17 vs 4/18, P= 0. 026). The same outcomes were obtained by quantitative Western blot images. Conclusion The intestinal epithelial permeability was increased in rats with obstructive jaundice,and intestinal barrier was damaged. After excising spleen, the amount and distribution of tight junction proteins were changed and the impairment of intestinal barrier was abated.