【作者】 郭佳汶； 程如越； 万群； 张瑜杰； 周玮忻； 王茂林； 李鸣； 何方；

【Author】 Jiawen Guo;Ruyue Cheng;Qun Wan;YuJie Zhang;Weixin Zhou;Maolin Wang;Ming Li;Fang He;West China School of Public Health,Sichuan University;

【机构】 四川大学华西公共卫生学院；

【摘要】 本实验旨在探究生命早期抗生素使用引起的肠道菌群紊对成年期免疫系统和过敏性疾病易感性的影响。54只BALB/C新生小鼠分为三组(N=18),出生后至21天,抗生素组每天灌胃万古霉素(100mg/kg),阴性对照组和阳性对照组用等量生理盐水,21天时每组处理6只,检测小鼠肠道绒毛、隐窝、肠上皮细胞相关蛋白表达,qPCR和二代测序分析粪便细菌总浓度及粪便细菌构成。21天停止干预后,万古霉素组和阳性对照组行腹腔OVA刺激,并继续饲养至56天,用同样方法观察受试小鼠成年期肠道上皮组织及粪便细菌组成,测定三组血清细胞因子TNFα、IL-4、IL-6、IL-10、IL-17A、IFN-γ水平及血清IgE含量并检测脾脏CD4+T细胞的比例。结果显示万古霉素干预21天后,与对照组相比,回肠和结肠绒毛变短变粗,排列松散。十二指肠、空肠、回肠绒毛高度和隐窝深度的比值(V/C)、十二指肠上皮细胞相关蛋白ki67、肠道菌群丰富度及多样性降低,而回肠、结肠ZO-1表达升高,另外肠道微生物构成也完全改变,变形菌门成为肠道优势菌。56天时肠道上皮和肠道菌群部分恢复,但空肠、回肠、结肠ZO-1的表达及个别菌群仍存在差异。投用万古霉素后血清细胞因子IL-6、IL-4/IFN含量较高,IL-17A含量较低。据此得出结论生命早期投用万古霉素能影响肠道菌群的构建,损伤机体肠道上皮细胞,对成人期过敏性疾病有一定的促进作用。更多还原

【Abstract】 This research aims to study how gut dysbacteriosis caused by antibotics in the early life alter the maturation of the immune system and the susceptibility of allergic disease in adulthood late. BALB/c neonatal mice were given by gavage for 3 weeks with vancomycin. Then the tested mice were immunized with intraperitoneal ovalbumin(OVA). Histopathological examinations determined the function and morphology of the host intestinal epithelium. Intestinal microbiota was detected and characterized by using quantitative polymerase chain reaction and next-generation sequencing of 16 S rRNA genes. Determinting the serum levels of tumor necrosis factor(TNF)-α, interleukin(IL)-6, IL-10, and IgE and the percentage of splenic CD4+ T cells by using enzyme-linked immunosorbent assay and flow cytometry, respectively. The results showed that at postnatal 21 day, compared with control group, vancomycin treatment resulted in shorter, thicker and looser villi of ileum and colon. The number of ki67–positive cells of duodenum and in ZO-1-positive cells of jejunum, colon in vancomycin treated mice were lower than the control. In addition, the composition of the intestinal flore between two groups were completely different from each other and Proteobacteria became the predominant bacteria. At postnatal day 56 after a period of discontinuing vancomycin treatment, intestinal epithelium and gut flora were partly recovered, but the expression of ZO-1 protein in jejunum, ileum, colon and specific intestinal flora remained different. Bacterioidetes was still significantly less in vancomycin treated mice, while Firmicutes was significantly increased. Additionally, serum IL-6 level and IL-4/IFN-γ value of vancomycin-treated group were significantly higher than control group while serum IL-17 A level in vancomycintreated group reduced. Our findings indicate that the oral administration of vancomycin during early life could injury intestinal mucosa and promote the risk of allergies in adult host animals. Alteration of gut flora composition may be an important mechanism underlying the impact of vancomycin on the immune sensitivity of hosts in adulthood late.更多还原