幽门螺杆菌感染性慢性胃炎模...区域的菌群分布特征及其机制_覃艳春.pdf

第 49 卷 第 2 期2023年 3 月吉林大学学报（医学版）Journal of Jilin University（Medicine Edition）Vol.49 No.2Mar.2023DOI：10.13481/j.1671587X.20230204幽门螺杆菌感染性慢性胃炎模型小鼠肠道各区域的菌群分布特征及其机制覃艳春1,黄衍强1,陆钢2,黄干荣1,唐华英1,戴园园1（1.右江民族医学院基础医学院病原生物学与免疫学教研室，广西 百色 533000；2.右江民族医学院附属医院烧伤整形与创面修复外科，广西 百色 533000）摘要 目的目的：探讨幽门螺杆菌（Hp）感染性慢性胃炎模型小鼠肠道各区域菌群种类、特征和菌群差异，并阐明其相关机制。方法方法：30只 C57BL/6小鼠随机分为对照组和感染组，每组 15只。对照组小鼠以生理盐水灌胃，感染组小鼠以浓度为 1109CFU mL1的 Hp 菌悬液灌胃，确认建模成功后采集 2 组小鼠的十二指肠、空肠和结肠内容物提取总 DNA，PCR 法扩增，采用高通量测序技术对样本 16SrRNAV3-V4区进行测序，通过 和 多样性分析对照组和感染组小鼠肠道菌群的特征、差异和多样性。结果结果：2 组样本聚类分析共产生的操作单元分类（OTU）数目为 211 个。在门水平，2 组样本主要有厚壁菌门、拟杆菌门、放线菌门、变形菌门和疣微菌门 5个优势菌门；与对照组比较，感染组小鼠十二指肠、空肠和结肠中的放线菌门丰度明显降低（P0.05）。在属水平，2组样本主要有乳酸菌属、葡萄球菌属、梭状芽孢杆菌属a 菌属、Turicibacter 菌属、阿克曼菌属、双歧杆菌属、Saccharibacteria-genera-incertae-sedis菌属、巴恩斯氏菌属、脱硫弧菌属、另枝菌属和支原体等 21个优势菌属；与对照组比较，感染组小鼠十二指肠和结肠中的乳酸菌属及双歧杆菌属丰度明显降低（P0.05），支原体丰度明显升高（P0.05）。Alpha多样性分析，shannon指数，对照组与感染组小鼠的十二指肠和空肠菌群多样性比较差异有统计学意义（P0.01），感染组菌群多样性降低；simpson指数，对照组与感染组小鼠的十二指肠、空肠和结肠菌群多样性比较差异有统计学意义（P0.05），感染组菌群多样性降低。Welch s t-test分析，2 组小鼠肠道 各 区 域 多 数 菌 群 丰 度 比 较 差 异 有 统 计 学 意 义（P0.05）。结论结论：Hp感染性慢性胃炎小鼠肠道各区域中菌群种类和特征发生改变，益生菌丰度降低，支原体丰度升高，其机制可能与 Hp的定植改变肠道微环境有关。关键词 幽门螺杆菌；胃炎；肠道菌群；小鼠，C57BL/6；高通量测序技术中图分类号 R378文献标志码 A文章编号 1671587X(2023)02028909收稿日期 20220627基金项目 国家自然科学基金项目（32060018）；广西壮族自治区中医药管理局自筹项目（GXZYZ20210135）；广西壮族自治区百色市科技局科学研究与技术开发计划项目（20211806）；广西壮族自治区百色市科技局百色市区域多发病联合专项计划项目（20224165）作者简介 覃艳春（1978），女，广西壮族自治区百色市人，高级实验师，主要从事医学微生物学和免疫学方面的研究。通信作者 唐华英，讲师（E-mail：）；陆 钢，副主任医师（E-mail：）289第 49 卷 第 2 期 2023 年 3 月吉林大学学报（医学版）Distribution characteristics of micrflora in various regions of intestinal tract of mice with chronic gastritis infected with Helicobacter Pylori and its mechannismQIN Yanchun1,HUANG Yanqiang1,LU Gang2,HUANG Ganrong1,TANG Huaying1,DAI Yuanyuan1（1.Department of Pathogenic Biology and Immunology，School of Basic Medical Sciences，Youjiang Medical University for Nationalities，Baise 533000，China；2.Department of Burn Plastic and Wound Repair Surgery，Affiliated Hospital，Youjiang Medical University for Nationalities，Baise 533000，China）ABSTRACT Objective：To investigate the types，characteristics，and differences of microflora in various regions of the intestinal tract of the mice with chronic gastritis infected with Helicobacter pylori（Hp），and to clarify the related mechanism.Methods：Thirty C57BL/6 mice were randomly divided into control group and infection group，and there were 15 mice in each group.The mice in control group were given normal saline，and the mice in infection group were given Hp suspension with the concentration of 1109 CFU mL1.After the model was successful builted，the contents of duodenum，jejunum，and colon of the mice in two groups were collected to extract the total DNA，and PCR amplification was performed；the 16SrRNAV3-V4 region was sequenced through high-throughput sequencing technique；the characteristics，differences，and diversities of the microflora in various regions of intestinal tract of the mice in control and infection group were analyzed by and diversity analysis.Results：The total number of operational taxonomic unit（OTU）in two groups generated by cluster analysis samples was 211.At the phylum level，there were 5 dominant bacteria such as Firmicutes，Bacteroidetes，Actinomyces，proteobacteria，and Verrucomicrobia；compared with control group，the abundances of Actinobacteria in the duodenum，jejunum and colon of the mice in infection group were significantly decreased（P0.05）.At the genus level，there were 21 dominant bacteria such as Lactobacillus，Staphylococcus，Clostridium Bacillus-a，Turicibacter，Akkermansia，Bifidobacterium，Saccharibacteria-genera-incertae-sedis，Barnesia，Desulfovibrio，Alternaria，and Mycoplasma and so on；compared with control group，the abundances of Lactobacillus and Bifidobacterium in the duodenum and colon of the mice in infection group were significantly decreased（P0.05），and the abundance of Mycoplasma was significantly increased（P0.05）.In the Alpha diversity analysis，the shannon index analysis results showed that there were significant differences in the diversities of duodenum and jejunum microflora of the mice between control group and infection group（P0.01），and the diversity of microflora in infection group was decreased；the simpson index analysis results showed that there were statistically significant differences in the diversities of duodenum，jejunum and colon microflora of the mice between control group and infection group（P0.05）；and the diversity of microflora in infection group was decreased.The Welch s t-test analysis results showed that the abundances in various regions of intestinal tract of the mice in two groups had signifricant differences（P0.05）.Conclusion：The types and characteristics of the microflora in various regions of intestinal tract of the mice with chronic gastritis infected with Hp change，the abundance of the probiotics is decreased and the abundance of Mycoplasma is increased.Its mechanism may be related with the changes of intestinal microenviroment，induced by the colonization of Hp.KEYWORDS Helicobacter