Page 33 - 南京医科大学学报自然科学版

第43卷第1期 南京医科大学学报（自然科学版）
2023年1月 Journal of Nanjing Medical University（Natural Sciences） · 27 ·

·基础研究·

益生菌A. muciniphila改善大肠杆菌诱导的小鼠腹泻

孙学伟 ，何君花 ，姜新泽 ，何世军 ，朱 磊 ，唐成亮 ，秦明珂 ，朱 进 ，杨 展 ，吴玉龙 1*
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滨州医学院基础医学院，山东 烟台 264003；东部战区总医院妇产科，江苏 南京 210002；陆军军医大学基础医学院，重
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庆 400038；东部战区疾病预防控制中心传染病防控二科，江苏 南京 210002
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［摘 要］ 目的：以产肠毒素大肠杆菌（enterotoxigenic Escherichia coli，ETEC）诱导肠道菌群失调性腹泻模型，分析益生菌嗜
黏蛋白阿克曼氏菌（Akkermansia muciniphila，A. muciniphila）对 ETEC 诱导腹泻的改善效果。方法：用 ETEC 诱导小鼠腹泻模
型。实验小鼠随机分为 PBS 组、模型组、干预组。干预组给予灌服 A. muciniphila 菌液（浓度为 1×10 CFU/mL）0.2 mL/d，连续
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5 d，模型组口腔灌服ETEC菌液（浓度为1×10 CFU/mL）0.5 mL/d。记录小鼠身体状况、体重、腹泻率和死亡率的变化；HE 染色
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观察结肠组织病理变化；ELISA 检测血清白细胞介素（interleukin，IL）IL⁃1β、IL⁃6、肿瘤坏死因子⁃α（tumor necrosis factor⁃α，
TNF⁃α）的含量；RT⁃PCR 检测结肠组织 IL⁃1、TNF⁃α、IL⁃6 mRNA 的表达；收取小鼠粪便进行 16S rRNA高通量测序并进行统计
分析。结果：干预组与模型组相比，身体状况明显改善，腹泻率降低了66%，死亡率降低了30%；血清IL⁃1β、IL⁃6和TNF⁃α含量
明显降低（P < 0.05）。结肠组织 IL⁃1、IL⁃6、TNF⁃α mRNA 含量明显降低（P < 0.05）。结肠组织病理变化显著改善。16S rRNA
测序显示干预组物种相对丰度明显增加，肠道菌群α多样性与菌群组成发生明显改变，在干预组中，多种益生菌丰度增加。
结论：益生菌A. muciniphila通过增加肠道菌群多样性、保护结肠黏膜组织和降低炎症因子的表达，减轻ETEC引起的腹泻。
［关键词］ 产肠毒素大肠杆菌；嗜黏蛋白阿克曼氏菌；腹泻；肠道菌群
［中图分类号］ R574 ［文献标志码］ A ［文章编号］ 1007⁃4368（2023）01⁃027⁃07
doi：10.7655/NYDXBNS20230104

Probiotics A. muciniphila improving diarrhea induced by ETEC in mice
SUN Xuewei ，HE Junhua ，JIANG Xinze ，HE Shijun ，ZHU Lei ，TANG Chengliang ，QIA Mingke ，ZHU Jin ，
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YANG Zhan ，WU Yulong 1*
1 Basic Medical College，Binzhou Medical College，Yantai 234003；Obstetrics and Gynecology Department，General
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Hospital of Eastern Theater Command of Chinese PLA，Nanjing 210002；Basic Medical College，Army Medical
University，Chongqing 400038；Infectious Disease Prevention and Control Division II，Eastern Theater Disease
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Prevention and Control Center，Nanjing 210002，China
［Abstract］ Objective：We used enterotoxigeni Escherichia coli（ETEC）to build the diarrhea model induced by intestinal flora
dysfunction. Then the probiotics Akkermansia muciniphila（A. muciniphila）was used to analyze the protective effect in the diarrhea.
Methods：The diarrhea model of mice was induced by ETEC. The experimental mice were randomly divided into PBS group，ETEC
model group and A. muciniphila treatment group. Each mouse of the A. muciniphila treatment group was treated with A. muciniphila
solution（1×10 CFU/mL）0.2 mL/d for 5 days，and the model group was gavaged with ETEC solution（concentration 1×10 CFU/mL）
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0.5 mL/d for 5 days. The changes of physical condition，body weight，diarrhea rate and mortality of mice were recorded；the pathological
changes of colon tissue were observed using HE staining；the contents of serum interleukin（IL）⁃1β，IL⁃6 and tumor necrosis factor⁃α
（TNF⁃α）were detected using ELISA method；the expression of IL⁃1，TNF⁃α and IL⁃6 mRNA in colon tissue were detected by RT⁃PCR；
and the feces of mice were collected for 16S rRNA high⁃throughput sequencing and statistical analysis. Results：Compared with the
ETEC model group，the physical condition of the A. muciniphila treatment group was significantly improved，the diarrhea rate was
reduced by 66% ，the mortality rate was reduced by 30% ，and the contents of serum IL ⁃ 1β，IL ⁃ 6 and TNF ⁃ α were significantly

［基金项目］ 国家生物安全专项（2019SWAQ05⁃5⁃4）；江苏省社会发展项目（BE2018617）
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通信作者（Corresponding author），E⁃mail：breezezhan@126.com；ylongwu@126.com