大肠杆菌亚型致脓毒症模型比较研究  

Comparative study on sepsis models induced by Escherichia coli subtypes

作　　者：张晶晶 孔宪斌 霍景瑞 王蕾[1] 刘颖[1] 杨晓晖[1] 田毅[1] 侯振江[1] 陈锋[3] 陈旭义[3] 孙世中[3] 夏天光 孙中磊 黄梦强 刘英富 ZHANG Jing-jing;KONG Xian-bin;HUO Jing-rui;WANG Lei;LIU Ying;YANG Xiao-hui;TIAN Yi;HOU Zhen-jiang;CHEN Feng;CHEN Xu-yi;SUN Shi-zhong;XIA Tian-guang;SUN Zhong-lei;HUANG Meng-qiangs;LIU Ying-fu(Science and Technology Experimental Center,Cangzhou Medical College,Cangzhou 061001,China;Graduate School of Tianjin University of Traditional Chinese Medicine;Department of Brain Center,the Affiliated Hospital of Armed Police Logistics College;Graduate School of Jinzhou Medical University;Graduate School of Tianjin Medical University)

机构地区：[1]沧州医学高等专科学校科技实验中心,061001 [2]天津中医药大学研究生院 [3]武警后勤学院附属医院脑科中心 [4]锦州医科大学研究生院 [5]天津医科大学研究生院

出　　处：《天津医药》2018年第6期585-589,共5页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(11672332、81772018); 天津市自然科学基金(17JCZDJC35400); 沧州市科学技术研究与发展指导计划(162302146、172302093)

摘　　要：目的探讨不同种类大肠杆菌诱导的脓毒症模型损伤程度。方法将152只小鼠随机数字表法分为对照组、DH5α组、44102组和25922组,每组38只。DH5α组、44102组和25922组分别腹腔注射300μL浓度为1.0×10~9CFU/kg的大肠杆菌DH5α、44102和25922,以不同种类大肠杆菌诱导脓毒症模型,对照组腹腔注射等体积生理盐水。(1)造模8 h后,每组各取4只小鼠,进行外周血细菌培养,计算细菌菌落数。(2)造模12 h后,通过酶联免疫吸附试验(ELISA)的方法检测血清中肿瘤坏死因子(TNF)-α、白介素(IL)-6水平(每组10只);采用蛋白免疫印迹法(Westernblot)测定血清中晚期炎症因子高迁移率族蛋白1(HMGB1)水平(每组4只);使用自动生化分析仪检测血清中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、尿素氮(BUN)、血肌酐(CR)水平(每组10只);取肺肝肾组织经甲醛固定后,进行苏木精-伊红(HE)染色(每组10只)。结果 DH5α组、44102组、25922组细菌菌落形成数量,炎症因子TNF-α、IL-6以及HMGB1蛋白,肝肾功能指标ALT、AST、CR、BUN均呈依次增高趋势(P<0.01)。DH5α组、44102组、25922组的肺泡结构损伤程度、肝炎性细胞浸润程度及肾小球萎缩程度由重到轻依次为25922组、44102组、DH5α组,对照组肺、肝、肾组织无明显损伤。结论大肠杆菌25922诱导脓毒症模型损伤严重,可用于研究脓毒症初期炎症风暴阶段的动物模型;大肠杆菌44102诱导脓毒症模型损伤适中,可用于无需限定脓毒症分期实验的动物模型;大肠杆菌DH5α诱导脓毒症模型损伤较轻,可用于探索脓毒症免疫抑制期治疗方法的动物模型。Objective To investigate the degrees of injury severity of sepsis models made by different kinds of Escherichia coli. Methods The 152 mice were randomly divided into control group, DH5α group, 44102 group, and 25922 group, with 38 rats in each group. DH5α group, 44102 group and 25922 group were intraperitoneally injected with 300 μL of Escherichia coli DH5o~, 44102 and 25922 at the concentration of 1.0 × 109 CFU/kg to prepare sepsis models of different kinds of Escherichia coli. Control group was injected intraperitoneally with the same amount of nomml saline. （1） After 8 h, four mice were taken from each group for peripheral blood bacterial culture. （2） After 12 h, ten mice in each group were used for measuring serum levels of TNF-o~ and IL-6 by enzyme-linked immunosorbent assay （ELISA）. （3） Western blot assay was used to determine the serum levels of high-mobility group protein （HMGB 1） in four mice of each group. （4） Ten mice in each group were used to measure serum levels of alanine transaminase （ALT）, aspartate aminotransferase （AST）, creatinine （CR） and blood urea nitrogen （BUN） by automatic biochemical analyzer. （5） After liver, lung and kidney tissues were fixed with formaldehyde, hematoxylin-eosin （HE） staining was performed （u=10 for each group）. Results In DH5o~ group, 44102 group and 25922 group, bacteria, ilfflammatoU cytokines TNF-o~, IL-6 and HMGB1 protein, liver and kidney indicators ALT, AST, CR and BUN showed a sequential increasing trend （P 〈 0.01）. The severe degrees of alveolar structure damage, hepatic cell infiltration and renal glomerular atrophy were DH5α group, 44102 group and 25922 group in turn. There were no obvious damages of lung, liver or kidney tissues in control group. Conclusion Escherichia coli 25922 induces severe sepsis injmT and can be used to study the animal models of the initial inflammatoU phase of sepsis. Escherichia coli 44102 induces moderate damage of sepsis and can be used in animal models that do not re

关 键 词：脓毒症 全身炎症反应综合征 疾病模型 动物 大肠杆菌 肿瘤坏死因子Α 白细胞介素6 

分 类 号：R321.5[医药卫生—人体解剖和组织胚胎学]