通腑理肺汤对脓毒症肠屏障损伤大鼠肠黏膜组织闭锁小带蛋白-1、闭合蛋白mRNA及蛋白表达的影响  

The effect of Tongfu Lifei decoction on the expression of ZO-1 and Occludin mRNA and protein in intestinal mucosa of sepsis rats with intestinal barrier injury

作　　者：柳蔓 吕波 李兰 陈立[3] LIU Man;LYU Bo;LI Lan;CHEN Li(Department of Pediatrics,Second Hospital Affiliated to Traditional Chinese Medicine College of Guizhou,Guiyang 550001,China)

机构地区：[1]贵州中医药大学第二附属医院儿科,贵州贵阳550001 [2]贵州中医药大学第一附属医院重症医学科,贵州贵阳550001 [3]广东省中医院急诊科,广东广州510120

出　　处：《陕西中医》2024年第1期8-12,共5页Shaanxi Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(81860846)。

摘　　要：目的:探讨通腑理肺汤(TFL)对脓毒症肠屏障损伤大鼠肠黏膜组织闭锁小带蛋白-1(ZO-1)、闭合蛋白(Occludin)mRNA及蛋白表达影响的实验研究。方法:随机抽取纯种SD大鼠40只,依次分为假手术组、模型组、TFL低剂量组和TFL高剂量组,共四组,每组10只。应用盲肠结扎穿孔(CLP)的造模方式制作脓毒症肠屏障损伤大鼠模型;假手术组只暴露盲肠,不做穿孔处理。TFL低剂量组造模成功后给予3.6 g/kg体重TFL灌胃;TFL高剂量组采用相同方法造模后,按7.2 g/kg体重TFL进行灌胃处理;模型组和假手术组按蒸馏水10 ml/kg体重灌胃,以上各组均每天干预1次,共7 d。7 d后处死各组大鼠,分离肠黏膜组织,采用RT-qPCR法检测肠黏膜组织ZO-1、Occludin mRNA基因,Western blot检测ZO-1、Occludin蛋白表达;同时取出各组相同部位的肠黏膜组织,光镜及透射电镜下观察肠黏膜组织病理及超微结构变化。结果:模型组大鼠肠黏膜组织中ZO-1、Occludin mRNA基因及蛋白表达对比假手术组均显著降低(P<0.05)。TFL低剂量组、高剂量组大鼠肠黏膜组织中ZO-1、Occludin mRNA基因及蛋白表达对比模型组均显著升高(P<0.05)。光镜下观察肠黏膜组织病理显示TFL低剂量组及高剂量组肠黏膜上皮细胞水肿、肠黏膜绒毛受损以及基底层、固有层水肿、淋巴细胞及中性粒细胞浸润明显减轻。透射电镜下肠黏膜组织超微结构亦显示TFL低剂量组及高剂量组微绒毛密集且规则,肠细胞间呈锯齿状和互锁模式,线粒体清晰,紧密连接轻微受损。结论:脓毒症肠屏障损伤大鼠模型病理损害的减轻,可能是TFL上调了肠黏膜组织紧密连接ZO-1、Occludin mRNA基因及蛋白的表达,改善肠上皮细胞间的紧密连接作用来实现的。Objective:To explore the effect of Tongfu Lifei decoction(TFL)on the mRNA genes and protein expression of ZO-1 and Occludin in intestinal mucosa tissue of sepsis rats with intestinal barrier injury.Methods:Randomly selected 40 purebred SD rats and divided them into four groups:Sham surgery group,model group,low dose TFL group,and high dose TFL group,with 10 rats in each group.Using the method of cecal ligation and perforation(CLP)to create a rat model of sepsis intestinal barrier injury.In the sham operation group,only the cecum was exposed without perforation.TFL low-dose group was given 3.6 g/kg body weight TFL by gavage after successful modeling.After modeling using the same method,the high-dose TFL group was treated with 7.2 g/kg body weight TFL by gavage.The model group and sham operation group were given 10 ml/kg of distilled water by gavage,and each group was intervened once a day for a total of 7 days.After 7 days,the rats in each group were killed,and the intestinal mucosa tissues were isolated.ZO-1 and Occludin mRNA genes were detected by RT-qPCR,and the expression of ZO-1 and Occludin protein was detected by Western blot.At the same time,took out the intestinal mucosa at the same part of each group,and observed the pathological and ultrastructural changes of intestinal mucosa under light and transmission electron microscope.Results:The mRNA genesand protein expressions of ZO-1 and Occludin in intestinal mucosal tissue of model group rats were significantly decreased compared with those of sham operation group(P<0.05).The mRNA genes and protein expressions of ZO-1 and Occludin in the intestinal mucosa of rats in the low-dose and high-dose TFL group were significantly increased compared to the model group(P<0.05).The histopathological observation of intestinal mucosa under light microscope showed that the edema of intestinal mucosal epithelial cells,the damage of intestinal mucosal villi,the edema of basal layer and lamina propria,the infiltration of lymphocytes and neutrophils were significantly reduced

关 键 词：闭锁小带蛋白-1 闭合蛋白 脓毒症肠屏障损伤 紧密连接蛋白 通腑理肺汤 超微结构 

分 类 号：R631.1[医药卫生—外科学]