摘要： 目的 探讨轮状病毒感染后乳鼠肠道菌群结构、小肠上皮细胞紧密连接蛋白及小肠通透性的改变。方法 构建轮状病毒感染乳鼠腹泻模型,收集结肠和直肠混合肠道内容物,利用16S rDNA测序技术检测乳鼠感染轮状病毒后肠道菌群结构及组成;采用实时荧光定量PCR检测轮状病毒感染后乳鼠粪便中病毒拷贝数及乳鼠肠道细胞紧密连接蛋白1(TJP-1)、紧密连接蛋白4(claudin-4)、链接黏附分子(F11R)基因的表达。采用FITC-葡聚糖检测乳鼠肠道上皮细胞的渗透性。结果 连续灌胃4 d后轮状病毒感染(RV)组乳鼠出现黄色水样粪便。16S rDNA测序结果显示,RV组与对照(NC)组的菌群分类组成有显著差异,筛选出10个显著差异菌属。NC组的优势菌属为Lactobacillus、Paenibacillus、Brevundimonas、Parabacteroides、Bacteroides和Alloprevotella,而RV组的富集菌属为Enterococcus和Shigella。实时荧光定量PCR结果显示,与NC组相比,RV组claudin-4及F11R基因表达水平显著升高(P<0.001),TJP-1表达水平显著降低(P<0.001)。RV组肠道通透性较NC组显著升高。结论 轮状病毒感染乳鼠后可能通过破坏肠道黏膜屏障的紧密连接蛋白改变肠道菌群的组成。

轮状病毒感染对乳鼠肠道菌群及小肠屏障功能的影响

陆恒章, 于美玲, 程美慧, 刘洋, 刘长城, 赵微

锦州医科大学基础医学院实验动物教研室, 辽宁 锦州 121000

收稿日期:2021-09-03出版日期:2022-12-30发布日期:2022-12-12
通讯作者:赵微E-mail:zhaowei-v@jzmu.edu.cn
作者简介:陆恒章 (1995-)，男，硕士研究生.
基金资助:辽宁省自然科学基金(2021-MS-334)；辽宁省教育厅青年育苗项目(JYTQN201925)


关键词: 轮状病毒, 肠道菌群, 紧密连接蛋白
Abstract: Objective To explore changes in the structure of the intestinal flora， tight junction proteins of small intestinal epithelial cells， and small intestinal permeability of suckling mice after rotavirus(RV) infection. Methods A RV infection model of diarrhea in suckling mice was constructed， and the mixed intestinal contents and tissues of the colon and rectum were collected. 16S rDNA sequencing was used to determine the structure and composition of the intestinal flora of suckling mice infected with rotavirus； real-time PCR was used to determine the virus copy number in stools of suckling mice after RV infection and expression of the genes coding for tight junction protein 1(TJP-1)， tight junction protein 4(claudin-4)， and link adhesion molecules(F11R). FITC-dextran was used to assay the permeability of the intestinal epithelial cells of neonatal rats. Results The suckling mice in the RV group produced yellow watery stools after four days of continuous gastric RV administration. The 16S rDNA sequencing results showed that there were significant differences in the taxonomic composition of the RV group versus that of the control(NC) group， and 10 significantly different bacterial genera were identified. The dominant bacteria genera in the NC group were Lactobacillus，Paenibacillus，Brevundimonas，Parabacteroides，Bacteroides， and Alloprevotella， while the enriched bacteria genera in the RV group were Enterococcus and Shigella. The real-time PCR Results showed that claudin-4 and F11R gene expression in the RV group were significantly higher than those in the NC group(P<0.001)， while TJP-1 gene expression was significantly reduced(P<0.001). The intestinal permeability of the RV group was significantly greater than that of the NC group. Conclusion Infection of suckling mice by rotavirus can change the composition of the intestinal flora by destroying tight junction proteins of the intestinal mucosal barrier.
Key words: rotavirus, intestinal flora, tight junction protein
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