严重烧伤小鼠肠-胰岛轴功能的变化及其作用  

Changes in entero-insular axis function and its role in mice with severe burns

作　　者：刘馨竹 李大伟[1] 蒋敏 李志生 冯柏塨 申传安[1] Liu Xinzhu;Li Dawei;Jiang Min;Li Zhisheng;Feng Baigong;Shen Chuanan(Senior Department of Burns and Plastic Surgery,the Fourth Medical Center of PLA General Hospital,Beijing 100048,China)

机构地区：[1]解放军总医院第四医学中心烧伤整形医学部,北京100048

出　　处：《中华烧伤与创面修复杂志》2024年第7期625-633,共9页Chinese Journal of Burns And Wounds

基　　金：国家自然科学基金面上项目(82072169、82272279);国家自然科学基金青年科学基金项目(82302799);解放军总医院青年自主创新科学基金项目(22QNFC011)。

摘　　要：目的探讨严重烧伤小鼠肠-胰岛轴功能的变化及其作用。方法该研究为实验研究。将90只雄性8~10周龄C57BL/6J小鼠按随机数字表法分为假伤组和烧伤组(每组45只小鼠),于烧伤组小鼠背部制备30%体表总面积Ⅲ度烫伤(以下称烧伤)创面,假伤组小鼠模拟致假伤。伤后24 h,检测空腹血糖(样本数为12)后,行腹腔糖耐量实验和口服糖耐量实验,并绘制血糖浓度-时间变化曲线,计算曲线下面积(样本数为6);分别于腹腔注射或灌胃给予葡萄糖溶液前及腹腔注射或灌胃给予葡萄糖溶液后30、60、120 min从心脏取血,采用酶联免疫吸附测定(ELISA)法检测血浆胰岛素和胰高血糖素样肽1(GLP-1)水平(样本数为3);取每组3只小鼠回肠组织,行免疫荧光染色及原位末端标记染色检测肠道L细胞GLP-1表达及凋亡水平;提取每组6只小鼠胰岛行葡萄糖刺激的胰岛素分泌实验,分别经低糖(2.8 mmol/L葡萄糖)和高糖(16.7 mmol/L葡萄糖)孵育后取上清液,采用ELISA法检测胰岛素水平。取36只雄性8~10周龄C57BL/6J小鼠,按随机数字表法分为假伤组、烧伤组和烧伤+艾塞那肽4(Ex-4)组(每组12只小鼠),对假伤组和烧伤组小鼠行同前对应处理,将烧伤+Ex-4组小鼠同烧伤组小鼠致伤后给予其GLP-1受体激动剂Ex-4。伤后24 h,提取小鼠胰岛,采用蛋白质印迹法检测重链结合蛋白(BIP)、蛋白激酶R样内质网激酶(PERK)、磷酸化PERK(p-PERK)、真核翻译起始因子2α(eIF2α)、磷酸化eIF2α(p-eIF2α)和CCAAT/增强子结合蛋白同源蛋白(CHOP)的蛋白表达并计算p-PERK/PERK、p-eIF2α/eIF2α比值(样本数为3),采用流式细胞术检测胰岛细胞凋亡率(样本数为3),同前行葡萄糖刺激的胰岛素分泌实验以检测上清液中胰岛素水平(样本数为6)。结果伤后24 h,烧伤组小鼠空腹血糖为(7.3±1.0)mmol/L,显著高于假伤组的(5.1±0.6)mmol/L(t=6.36,P<0.05)。伤后24 h,在腹腔糖耐量实验和口服糖耐量实验中,烧�Objective To explore the changes in entero-insular axis function and its role in mice with severe burns.Methods This study was an experimental study.Ninety C57BL/6J male mice aged 8-10 weeks were divided into sham injury group and burn group(with 45 mice in each group)according to the random number table.A full-thickness scald(hereinafter referred to as burn)wound of 30%of the total body surface area was created on the back of mice in burn group,and the mice in sham injury group were simulated to cause a sham injury.Twenty-four hours after injury,the fasting blood glucose was measured(n=12),followed by intraperitoneal glucose tolerance test and oral glucose tolerance test;the curve of blood glucose concentration changes over time was plotted,and the area under the curve was calculated(n=6);the blood was taken from the heart before intraperitoneal injection or gavage of glucose solution and at 30,60,and 120 minutes after intraperitoneal injection or gavage of glucose solution for measuring the plasma insulin and glucagon like peptide-1(GLP-1)levels using enzyme-linked immunosorbent assay(ELISA),with a sample number of 3;the ileal tissue was taken from 3 mice in each group for detecting the GLP-1 expression and apoptosis levels of intestinal L cells by immunofluorescence staining and TdT-mediated dUTP nick-end labeling staining;the pancreatic islets were collected from 6 mice in each group for glucose-stimulated insulin secretion experiments.After incubation with low glucose(2.8 mmol/L glucose)and high glucose(16.7 mmol/L glucose),the supernatant was taken and the insulin level was detected using ELISA.Thirty-six C57BL/6J male mice aged 8-10 weeks were divided into sham injury group,burn group,and burn+exendin-4(Ex-4)group(with 12 mice in each group)according to the random number table.The mice in sham injury group and burn group were subjected to the same corresponding treatment as before.The mice in burn+Ex-4 group were injured in the same way as the burn group mice followed by treatment with GLP-1 receptor agoni

关 键 词：烧伤 胰岛素 胰高血糖素样肽1 内质网应激 高糖血症 肠-胰岛轴 

分 类 号：R644[医药卫生—外科学]