肝硬化患者肠道菌群损伤小鼠肠黏膜屏障促肝硬化发展  

Gut microbiota of patients with cirrhosis promotes cirrhosis development by injuring intestinal mucosal barrier in mice

作　　者：赵阳[1] 郭瑛[1] 黄娜[1] 陈曦[1] 张艾煜 柳蕾 孔光耀 杨军[2] 李宗芳[1] Zhao Yang;Guo Ying;Huang Na;Chen Xi;Zhang Aiyu;Liu Lei;Kong Guangyao;Yang Jun;Li Zongfang(National&Local Joint Engineering Research Center of Biodiagnosis and Biotherapy,the Second Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710004,China;Department of Pathology,the Second Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710004,China)

机构地区：[1]西安交通大学第二附属医院生物诊断治疗国家地方联合工程研究中心,710004 [2]西安交通大学第二附属医院病理科,710004

出　　处：《中华实验外科杂志》2021年第8期1567-1570,共4页Chinese Journal of Experimental Surgery

基　　金：陕西省自然科学基础研究计划青年人才项目(2018JQ8052);西安交通大学第二附属医院科研基金项目(2020YJ(ZYTS)335)。

摘　　要：目的观察乙肝肝硬化患者肠道菌群对肝硬化小鼠肠黏膜屏障功能及肝硬化进程的影响。方法收集西安交通大学第二附属医院2017年9月至2018年6月入院的乙肝肝硬化患者和健康人的粪便样品各12例,16S rDNA测序分析肠道菌群结构并制备菌液。雄性C57BL/6小鼠采用随机数表法分为4组:正常对照组(HC组)、肝硬化模型组(LC组)、模型组+健康人菌液(FMT-HC组)、模型组+患者菌液(FMT-LC组)。造模同时用菌液灌胃处理小鼠,持续12周。苏木精-伊红染色观察结肠病理形态,酶联免疫吸附试验和荧光探针检测肠黏膜屏障损伤,实时定量聚合酶链反应和免疫荧光检测肠黏膜屏障和肝纤维化相关因子的表达。两组间比较采用t检验,多组间比较采用One-way ANOVA分析。结果FMT-LC组小鼠的结肠中黏蛋白-2和闭合蛋白-2的表达量低于FMT-HC组(0.36±0.21比1.00±0.36,t=4.659,P<0.01和0.61±0.42比1.00±0.17,t=2.596,P<0.05),外周血中异硫氰酸荧光素-葡聚糖和脂多糖的浓度高于FMT-HC组[(9.32±2.24)μg/ml比(5.23±1.34)μg/ml,F=15.118,P<0.01和(88.55±10.50)ng/ml比(76.48±10.13)ng/ml,F=5.473,P<0.05],差异均有统计学意义。FMT-LC组小鼠的肝脏中转化生长因子-β1(3.39±2.67比0.95±0.12,t=2.744,P<0.05)等多种促纤维化因子和α-平滑肌肌动蛋白的表达量高于FMT-HC组,肝损伤加重。结论乙肝肝硬化患者肠道菌群降低肝硬化小鼠肠黏膜屏障功能,促肝硬化发展。Objective To investigate the effects of gut microbiota in patients with hepatitis B cirrhosis on function of intestinal mucosal barrier and progression of cirrhosis in mice.Methods Fecal samples of 12 patients and 12 healthy individuals in the Second Affiliated Hospital of Xi′an Jiaotong University from September 2017 to June 2018 were collected.Gut microbiota structure was analyzed through 16S rDNA sequencing and bacterial suspension was prepared.Male C57BL/6 mice were randomly(random number)divided into four groups:healthy control group(HC group),cirrhosis model group(LC group),model group+bacterial suspension of healthy individuals(FMT-HC group),model group+bacterial suspension of patients(FMT-LC group).Cirrhosis model was established and mice were gavaged with bacterial suspension,lasting for 12 weeks.Colon pathological morphology was observed by hematoxylin-eosin staining,injury of intestinal mucosal barrier was detected using enzyme-linked immunosorbent assay and fluorescent probe,the expression of intestinal mucosal barrier and hepatic fibrosis related factors were detected by real-time quantitative polymerase chain reaction and immunofluorescence.Results The expression levels of mucin-2 and claudins-2 in the colon of mice in FMT-LC group were significantly lower than those in FMT-HC group(0.36±0.21 vs.1.00±0.36,t=4.659,P<0.01 and 0.61±0.42 vs.1.00±0.17,t=2.596,P<0.05),and the concentrations of FITC-dextran and lipopolysaccharide in the peripheral blood were significantly higher than those in FMT-HC group[(9.32±2.24)μg/ml vs.(5.23±1.34)μg/ml,F=15.118,P<0.01 and(88.55±10.50)ng/ml vs.(76.48±10.13)ng/ml,F=5.473,P<0.05].The expression levels of transforming growth factor-β1(3.39±2.67 vs.0.95±0.12,t=2.744,P<0.05)and other pro-fibrotic factors,as well asα-smooth muscle actin in the liver of mice in FMT-LC group were significantly higher than those in FMT-HC group,and the liver injury was aggravated.Conclusion The gut microbiota of patients with hepatitis B cirrhosis can reduce the intestinal muco

关 键 词：肝硬化 肠道菌群 DNA测序 肠黏膜屏障 

分 类 号：R575.2[医药卫生—消化系统]