粪菌移植对严重烧伤大鼠肠道屏障功能的作用及相关机制  

Effect and related mechanism of faecal microbiota transplantation on intestinal barrier function in severely burned rats

作　　者：陈强 曹胜军[1] 巴特[1] 赵翠娟 Chen Qiang;Cao Shengjun;Ba Te;Zhao Cuijuan(Department of Burn,Third Affiliated Hospital of Inner Mongolia Medical University(Inner Mongolia Baogang Hospital),Baotou 014010,China;Department of Gastrorenterology,Third Affiliated Hospital of Inner Mongolia Medical University(Inner Mongolia Baogang Hospital),Baotou 014010,China)

机构地区：[1]内蒙古医科大学第三附属医院(内蒙古包钢医院)烧伤科,包头014010 [2]内蒙古医科大学第三附属医院(内蒙古包钢医院)消化内科,包头014010

出　　处：《中华损伤与修复杂志（电子版）》2022年第4期322-329,共8页Chinese Journal of Injury Repair and Wound Healing(Electronic Edition)

基　　金：中国金属学会冶金安全与健康分会健康卫生科研项目(jkws201833);内蒙古自治区自然科学基金项目(2020MS08024)。

摘　　要：目的探讨粪菌移植(FMT)对严重烧伤大鼠肠道屏障功能的作用及相关机制。方法按照随机数字表法将36只6~8周龄雄性SD大鼠分为3组:正常组(n=12)、单纯烧伤组(n=12)和FMT干预组(n=12)。单纯烧伤组和FMT干预组大鼠制作30%总体表面积Ⅲ度烫伤模型;正常组大鼠不致伤。单纯烧伤组、FMT干预组大鼠伤后即刻腹腔注射平衡盐溶液40 mL/kg补液复苏。收集正常组大鼠新鲜粪便10 g,制成粪便滤液。伤后1 h,对FMT干预组大鼠进行粪便滤液灌胃(10 mL/kg),间隔12 h后再次给予相同剂量粪便滤液灌胃;正常组、单纯烧伤组大鼠在相同时相点均给予等量0.9%氯化钠溶液灌胃。分别于伤后24、72 h,取伤后各组大鼠结肠内新鲜粪便各1 mL,采用实时荧光定量-聚合酶链反应检测大鼠肠道菌群属水平,即双歧杆菌、脆弱拟杆菌、乳酸杆菌、大肠杆菌、肠球菌数量;取制备好的大鼠血浆,采用荧光素异硫氰酸酯(FITC)-葡聚糖通透性实验检测3组大鼠肠道通透性;采用酶联免疫吸附试验(ELISA)检测大鼠血清中二胺氧化酶、D-乳酸及白细胞介素(IL)-6、肿瘤坏死因子(TNF)-α、IL-10水平;分别取伤后24、72 h 3组大鼠的末端回肠组织约1 cm,苏木精-伊红染色观察大鼠肠黏膜组织形态结构。数据比较采用单因素方差分析和t检验。结果(1)3组大鼠在伤后24、72 h双歧杆菌、脆弱拟杆菌、乳酸杆菌、大肠杆菌、肠球菌数量比较,差异均有统计学意义(P<0.05)。伤后24 h,单纯烧伤组大鼠肠道双歧杆菌、脆弱拟杆菌、乳酸杆菌、大肠杆菌、肠球菌数量分别为(2.76±0.15)、(3.27±0.40)、(2.33±0.33)、(7.06±0.49)、(6.42±0.50)LogN/g,FMT干预组大鼠分别为(3.18±0.16)、(4.52±0.58)、(2.92±0.28)、(6.14±0.47)、(5.28±0.43)LogN/g;伤后72 h,单纯烧伤组大鼠肠道双歧杆菌、脆弱拟杆菌、乳酸杆菌、大肠杆菌、肠球菌数量分别为(3.16±0.19)、(3.79±0.42)、(2.64±0.43)、(6.34±0.56)Objective To investigate the effect and related mechanism of faecal microbiota transplantation(FMT)on intestinal barrier function in severely burned rats.Methods Thirty-six male SD rats aged 6-8 weeks were divided into 3 groups according to the random number table method,normal group(n=12),pure burn group(n=12)and FMT intervention group(n=12).Thirty percent total body surface area full thickness burn model was made for rats in pure burn group and FMT intervation group,rats in normal group were not injured.Rats in pure burn group and FMT intervention group were resuscitated by intraperitoneal injection of balanced salt solution 40 mL/kg immediately after burn.Fresh faecal samples of the normal group rats were collected 10 g and made into faecal filtrate.At 1 h after injury,rats in the FMT intervention group was given faecal filtrate by gavage(10 mL/kg),and the same dose of faecal filtrate was given again 12 h later.Rats in normal group and pure burn group were given the same amount of 0.9%sodium chloride solution at the same time.At 24 and 72 h after injury,1 mL fresh faecal samples from the colon of rats in each group were collected,and the level of intestinal flora was detected by real-time fluorescence quantification-polymerase chain reaction,namely,the number of Bifidobacteria,Bacteroides fragilis,Lactobacillus,Escherichia coli and Enterococcus.The prepared plasma of rats was taken,and luciferin isothiocyanate(FITC)-glucan permeability test was used to detect the intestinal permeability of rats in each group.The levels of diamine oxidase,D-lactic acid,interleukin(IL)-6,tumor necrosis factor(TNF)-αand IL-10 in serum were detected by enzyme-linked immunosorbent assay(ELISA).At 24 and 72 h after injury,about 1 cm of the terminal ileum tissue of rats in each group were taken respectively,and the morphological structure of intestinal mucosa was observed by hematoxylin-eosin staining.Data were processed by one-way ANOVA and t test.Results(1)The differences in the number of Bifidobacteria,Bacteroides fragilis,Lactob

关 键 词：烧伤 大鼠 肠道菌群 肠道屏障 粪菌移植 

分 类 号：R644[医药卫生—外科学] R574[医药卫生—临床医学]