动物营养学报 >

2019 , Vol. 31 >Issue 3: 1278 - 1287

DOI: https://doi.org/10.3969/j.issn.1006-267x.2019.03.033

分子营养 Molecular Nutrition

抗生素诱导肠道菌群失调对肠黏膜屏障和肝脏功能的动态影响

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  • 1. 广州中医药大学, 广州 510006;
    2. 邓州市职业技术学校, 邓州 474150
李娜(1994-),女,安徽宿州人,硕士研究生,研究方向为中药药效评价与作用机制。E-mail:Lee-Nagzucm@foxmail.com

收稿日期: 2018-08-31

  网络出版日期: 2019-03-18

基金资助

国家自然科学基金项目（81202997）；广东省科技计划项目（2015A030302071）；广州市科技计划项目（201607010333）

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Dynamic Effects of Antibiotic-Induced Intestinal Dysbacteriosis on Intestinal Mucosa Barrier and Liver Function

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  • 1. Guangzhou University of Chinese Medicine, Guangzhou 510006, China;
    2. Dengzhou Vocational and Technical College, Dengzhou 474150, China

Received date: 2018-08-31

  Online published: 2019-03-18

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摘要

为了观察不同程度肠道菌群失调模型中肠黏膜屏障和肝脏功能的动态变化，采用头孢曲松钠复制小鼠肠道菌群失调模型，分别于造模处理的第3天、第6天、第10天取小鼠盲肠内容物进行16S rDNA基因测定，苏木精-伊红（HE）染色观察回肠组织病理变化，透射电镜观察回肠组织紧密连接情况，免疫组织化学法观察回肠组织闭锁小带蛋白-1（ZO-1）的阳性表达情况，酶联免疫吸附测定（ELISA）法检测肠组织匀浆中白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）的含量，同时检测肝脏功能指标，包括肝组织匀浆中谷丙转氨酶（ALT）、谷草转氨酶（AST）的活性及肝组织病理变化。结果显示：与正常组相比，头孢曲松钠处理3、6或10 d后，模型组小鼠肠道中除金黄杆菌属、普氏菌属、假单胞菌属的相对丰度逐步上升外，芽孢杆菌属、Lotus、葡萄球菌属的相对丰度逐步下降。模型组小鼠回肠组织中ZO-1阳性表达和肠上皮细胞间的紧密连接随着头孢曲松钠处理时间的延长逐渐下降。与正常组相比，模型组肠组织匀浆中TNF-α、IL-1β含量在菌群失调初期（头孢曲松钠处理3、6 d时）无显著变化（P>0.05），菌群持续失调（头孢曲松钠处理10 d时）后其含量极显著上升（P<0.01）；同时，菌群持续失调会累及肝脏，表现为模型组肝组织匀浆中ALT、AST活性较正常组极显著上升（P<0.01），肝细胞肿胀，肝索结构紊乱。以上结果提示，长时间的肠道菌群失调会导致肠道稳态被破坏，表现为肠黏膜屏障的破坏加重，肠道中炎症因子的含量逐渐上升，同时肝脏功能受到影响。

关键词： 头孢曲松钠; 肠道菌群失调; 肠黏膜屏障; 紧密连接; 肝脏损伤

本文引用格式

李娜, 赵玉民, 陈正元, 王碧君, 罗霞, 王青, 何国林, 熊天琴, 桑传兰 . 抗生素诱导肠道菌群失调对肠黏膜屏障和肝脏功能的动态影响[J]. 动物营养学报, 2019 , 31(3) : 1278 -1287 . DOI: 10.3969/j.issn.1006-267x.2019.03.033

Abstract

In order to observe the dynamic changes of intestinal mucosa barrier and liver function for different degrees of intestinal dysbacteriosis models, mouse models were made which had intestinal dysbacteriosis caused by ceftriaxone sodium. After administration of ceftriaxone sodium for 3, 6 and 10 days, the intestinal contents were sampled to measure 16S rDNA gene, the histopathological changes of the ileum tissues were observed by hematoxylin-eosin (HE) staining, the tight junction conditions of ileum tissues were observed by transmission electron microscope (TEM), the positive expression condition of zonula occluden-1 (ZO-1) of ileum tissues was observed by immunohistochemical (IHC) method, the contents of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in in intestinal homogenate were assessed by enzyme-linked immunosorbent assay (ELISA) method, the liver function indexes such as alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in liver homogenate and histopathological changes of liver tissue were also evaluated. The results showed as follows:compared with the normal group, the relative abundances of Chryseobacterium, Prevotella and Pseudomonas increased gradually, and the relative abundances of Bacillus, Lotus and Staphylococcus gradually decreased in model group after administration of ceftriaxone sodium for 3, 6 or 10 days. The positive expression of ZO-1 in ileum tissues was gradually decreased and the tight junction among the intestinal villus epithelial cells was gradually disappeared with the extension of ceftriaxone sodium treatment. Compared with the normal group, the contents of TNF-α and IL-1β in intestinal homogenate of model group had no changes at pre-dysbacteriosis (after administration of ceftriaxone sodium for 3 and 6 days), but increased significantly after the continued dysbacteriosis (after administration of ceftriaxone sodium for 10 days) (P<0.01). Meanwhile, with the ongoing imbalance of intestinal flora, it could affect the liver, which showed the activities of ALT and AST in liver homogenate in model group were significantly higher than those in normal group (P<0.01), and hepatocyte swelling and disordered structure of hepatic lobule were obviously found. It is concluded that long-term intestinal dysbacteriosis can destroy the intestinal microecological balance, which is reflected in the contents of intestinal proinflammatory cytokines are gradually increased, and the liver function is also affected.

Key words： ceftriaxone sodium; intestinal dysbacteriosis; intestinal mucosa barrier; tight junction; liver damage

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