肿瘤坏死因子相关凋亡诱导配体基因敲除对葡聚糖硫酸钠诱导结肠炎小鼠肠道菌群的影响  

Effect of tumor necrosis factor-related apoptosis-inducing ligand gene knock-out on the gut microbiota in mice with dextran sodium sulphate induced colitis

作　　者：郑淑姿 黄开宇[1] 吴昊[2] 蔡静 叶方鹏[4] 蒋益[2] Zheng Shuzi;Huang Kaiyu;Wu Hao;Cai Jing;Ye Fangpeng;Jiang Yi(Department of Pediatric Gastroenterology,The Second Affiliated Hospital and Yuying Children′s Hospital of Wenzhou Medical University,Wenzhou 325000,China;Department of Gastroenterology,The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University,Wenzhou 325000,China;Department of Gastroenterology,The Wenzhou Central Hospital,Wenzhou 325000,China;Department of Gastroenterology,Ruian People′s Hospital,Wenzhou 325200,China)

机构地区：[1]温州医科大学附属第二医院,育英儿童医院儿童消化科,325000 [2]温州医科大学附属第二医院,育英儿童医院消化内科,325000 [3]温州市中心医院消化内科,325000 [4]瑞安市人民医院消化内科,浙江省温州市325200

出　　处：《中华消化杂志》2019年第11期759-764,共6页Chinese Journal of Digestion

基　　金：浙江省自然科学基金(LY14H030012、LY15H030018、LY16H160055、LY17H030011);温州市科技局资助项目(Y20160102、Y20170062、Y20170314)。

摘　　要：目的在肿瘤坏死因子相关凋亡诱导配体基因敲除(TRAIL-/-)小鼠中诱导实验性结肠炎模型,分析肿瘤坏死因子相关凋亡诱导配体(TRAIL)缺乏对小鼠结肠炎症和肠道菌群组分的影响。方法选取C57BL/6品系的TRAIL-/-小鼠和野生型小鼠各24只,分为TRAIL-/-对照组(8只)和TRAIL-/-结肠炎组(16只),以及野生型对照组(8只)和野生型结肠炎组(16只)。两组结肠炎小鼠均连续7 d口服3.5%葡聚糖硫酸钠饮用水,诱导实验性结肠炎模型,从临床表现和组织病理学方面评估结肠炎严重程度。收集小鼠结肠组织标本,采用高通量16S rDNA测序技术检测结肠菌群组分,再应用USEARCH软件和R语言软件分析TRAIL-/-对照组、TRAIL-/-结肠炎组、野生型对照组和野生型结肠炎组小鼠结肠菌群组分的差异。组间比较采用独立样本t检验和秩和检验。结果造模开始后,野生型结肠炎组小鼠和TRAIL-/-结肠炎组小鼠的结肠炎疾病活动指数(DAI)都随时间推移逐渐增高,TRAIL-/-结肠炎组小鼠比野生型结肠炎组小鼠更早出现体质量下降、腹泻和便血。在造模后第7天,TRAIL-/-结肠炎组小鼠体质量下降(28.98±2.84)%,野生型结肠炎组小鼠体质量下降(17.87±3.70)%,两组间比较差异有统计学意义(t=9.53,P<0.01);TRAIL-/-结肠炎组小鼠的结肠长度短于野生型结肠炎组小鼠[(4.63±0.28)cm比(6.02±0.41)cm],差异有统计学意义(t=11.20,P<0.01);TRAIL-/-结肠炎组小鼠的DAI高于野生型结肠炎组小鼠(3.00±0.00比2.32±0.05),差异有统计学意义(t=54.40,P<0.01);TRAIL-/-结肠炎组小鼠的组织学评分高于野生型结肠炎组小鼠[(6.19±0.25)分比(3.87±0.22)分],差异有统计学意义(t=27.87,P<0.01)。与野生型结肠炎组小鼠相比,TRAIL-/-结肠炎组小鼠的结肠黏膜上皮受损、隐窝结构破坏和炎症细胞浸润更加明显。与野生型结肠炎组小鼠相比,TRAIL-/-结肠炎组小鼠的肠道菌群具有更高的α多样性:在科分类水平,TObjective To investigate the influence of tumor necrosis factor-related apoptosis-inducing ligant(TRAIL)deficiency on mice colitis and the gut microbiota composition by inclding the expermental colitis model in tumor necrosis factor-related apoptosis-inducing ligand gene knockout(TRAIL-/-)mice.Methods C57BL/6 TRAIL-/-mice and wild type(WT)mice were selected and assigned into TRAIL-/-control group(eight mice),TRAIL-/-colitis group(16 mice),WT control group(eight mice)and WT colitis group(16 mice).The mice of two colitis groups were oral administrated with 3.5%dextran sulphate sodium(DSS)in drinking water for seven consecutive days to induce experimental colitis model.The severity of colitis was evaluated by clinical appearance and histopathological examination.The colonic tissue samples of mice were collected and microbiota profile was analyzed by 16S rDNA sequencing method.USEARCH software and R language were used to analyze the difference of gut microbiota among TRAIL-/-control group,TRAIL-/-colitis group,WT control group and WT colitis group.T test and Mann-Whitney U test were used for statistical analysis.Results After modeling,the disease activity index(DAI)of WT colitis mice and TRAIL-/-colitis mice both gradually increased over time.Furthermore,compared with colitis mice,TRAIL-/-colitis mice developed body weight loss,diarrhea and hemafecia earlier.On the seventh day after modeling,the percentage of body weight loss of TRAIL-/-colitis mice and WT colitis mice was(28.98±2.84)%and(17.87±3.70)%,respectively;and the difference was statistically significant(t=9.53,P<0.01).The length of colon of TRAIL-/-colitis mice was shorter than that of WT colitis mice((4.63±0.28)cm vs.(6.02±0.41)cm),and the difference was statistically significant(t=11.20,P<0.01).The DAI of TRAIL-/-colitis mice was higher than that of WT colitis mice(3.00±0.00 vs.2.32±0.05),and the difference was statistically significant(t=54.40,P<0.01).The histological score of TRAIL-/-colitis mice was higher than that of WT colitis mice(6.19±0.25 v

关 键 词：TNF相关凋亡诱导配体 肠道菌群 实验性结肠炎 小鼠 基因敲除 

分 类 号：R57[医药卫生—消化系统]