甘油可能通过水通道蛋白3减轻脓毒症小鼠肠黏膜屏障损伤  

Glycerol may reduce intestinal mucosal barrier damage in sepsis mice through aquaporin 3

作　　者：何锐 朱烨柯 滕文彬 单跃 易声华[2] 祝胜美[1] 李玉红[2,3] HE Rui;ZHU Ye-ke;TENG Wen-bin;SHAN Yue;YI Sheng-hua;ZHU Sheng-mei;LI Yu-hong(Department of Anesthesiology, the First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310006;Department of Anesthesiology;Clinical Research Center, Shaoxing People's Hospital, Shaoxing 312000, China)

机构地区：[1]浙江大学医学院附属第一医院麻醉科,浙江杭州310006 [2]绍兴市人民医院麻醉科,浙江绍兴312000 [3]绍兴市人民医院医学研究中心,浙江绍兴312000

出　　处：《基础医学与临床》2020年第5期655-661,共7页Basic and Clinical Medicine

基　　金：浙江省科学技术厅公益项目(LGF19H030011);浙江省卫计委医药卫生科技计划(2018KY173,2019306157);绍兴市麻醉学重点学科(2019SZD04)。

摘　　要：目的探讨甘油对脓毒症小鼠肠黏膜屏障功能的保护作用。方法将小鼠随机分为4组:假手术组、假手术+口服甘油组、脓毒症模型组[小鼠(n=30),用盲肠结扎穿孔术(CLP)建立脓毒症模型,其中24只分别于建模后0、6、12和24 h处死,每个时点6只,作为时间依赖实验(时间依赖)]、脓毒症+口服甘油组。除时间依赖组,4组小鼠建模后24 h处死,收集血液和肠黏膜组织标本。用HE染色观察小肠黏膜病理损伤;ELISA测定二胺氧化酶(DAO)和肠型脂肪酸结合蛋白(FABP2)血浆浓度,检测肠黏膜通透性;免疫组化、RT-qPCR、Western blot检测肠黏膜水通道蛋白3(AQP3)表达。结果脓毒症诱导肠黏膜形态损伤,形态无序破坏,Chiu's评分呈时间依赖性升高(P<0.05);血浆DAO和FABP2浓度显著升高(P<0.05);AQP3表达随肠黏膜损伤加重表达明显下调(P<0.05)。口服甘油治疗部分改善脓毒症诱导的肠黏膜损伤,表现为肠黏膜形态结构改善、Chiu's评分降低以及血浆DAO和FABP2浓度降低。结论口服甘油治疗可能通过AQP3减轻脓毒症小鼠肠黏膜屏障损伤。其具体机制有待于进一步研究。Objective To explore the protective effect of oral glycerol on sepsis induced intestinal mucosal barrier function.Methods Mice were randomly divided to four groups:sham operation(n=6);sham operation with oral glycerol(n=6);Mouse sepsis model(n=30)was established using cecal ligation and puncture(CLP),of which 24 mice were executed at 0,6,12 and 24 h after CLP respectively(6 mice at each time point)as a time-dependent group,and sepsis with oral glycerol group(n=6).Besides the time-dependent group,four groups of mice were executed at 24 h after modeling.Blood and intestinal mucosal tissue samples were collected.HE staining was used to detect pathological damage of intestinal mucosa.The plasma diamine oxidase(DAO)concentration and intestinal-type fatty acid-binding protein 2(FABP2)concentration were measured by ELISA to assess intestinal mucosal permeability.Immunohistochemistry,real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and Western blot were used to detect the expression of AQP3 in intestinal mucosa.Results Sepsis induced morphological damage of intestinal mucosa,and the Chiu's score increased in a time-dependent manner(P<0.05).The concentrations of DAO and FABP2 in plasma were significantly increased(P<0.05),and the expression of AQP3 significantly down-regulated with the aggravation of injury(P<0.05).The administration of oral glycerol partially improved the sepsis-induced injury of the intestinal mucosa,as shown by partial recovery of the morphological structure,with decreased Chiu's score,decreased plasma concentrations of DAO and FABP2(sepsis vs sepsis+glycerol,P<0.05).Conclusions Oral treatment with glycerol may alleviate intestinal mucosal barrier damage in sepsis mice through aquaporin-3.The specific mechanism needs to be further studied.

关 键 词：脓毒症 肠道损伤 水通道蛋白3 甘油 

分 类 号：R363[医药卫生—病理学]