妊娠期母体使用抗生素和益生菌对后代小鼠肠道发育和肠道菌群的影响

EFFECTS OF MATERNAL USE OF ANTIBIOTICS AND PROBIOTICS DURING PREGNANCY ON INTESTINAL DEVELOPMENT AND FLORA OF OFFSPRING IN MICE

目的 探究妊娠晚期使用头孢曲松和两歧双歧杆菌TMC3115对断乳期小鼠肠道上皮组织发育及肠道菌群构建的影响。方法 24只妊娠10~13 d BALB/C小鼠随机分为对照组、头孢曲松组和TMC3115组(n=8)。妊娠期小鼠分别每日用生理盐水、头孢曲松(30 mg/d)、TMC31115菌悬液(含菌量为109 CFU/d)灌胃至分娩,新生小鼠由母鼠母乳喂养至断乳期(PND21)。采用HE染色观察小鼠结肠绒毛和隐窝深度;RT-PCR法检测各组小鼠肠道上皮组织发育相关蛋白(Ki67、Muc2)和紧密连接蛋白(ZO-1、Claudin-1、Claudin-2、Occludin)表达;二代测序法分析小鼠肠道菌群组成。结果 和对照组和头孢曲松组相比,TMC3115组小鼠的结肠隐窝深度、KI67 mRNA表达量显著(P<0.05)升高。TMC3115组小鼠紧密连接蛋白Occludin mRNA表达量也显著高于对照组(P<0.05)。对照组和TMC3115组肠道菌群中厚壁菌门相对丰度,头孢曲松组拟杆菌门相对丰度最高。和头孢曲松组相比,TMC3115组厚壁菌门和乳杆菌属相对丰度显著升高(P<0.05),梭菌属和肠球菌属相对丰度明显减少(P<0.05)。结论 妊娠期头孢曲松干预对子代肠道发育无明显影响,但能显著破坏子代肠道菌群结构,以潜在致病菌肠球菌属、梭菌属等增加为特征;而TMC3115干预能显著增加肠道厚壁菌门和乳杆菌属并促进肠道组织发育和屏障功能。

Objective To explore the effects of ceftriaxone and Bifidobacterium bifidum TMC3115 on the development of intestinal epithelial tissue and the establishment of intestinal flora of weaning offspring mice in late pregnancy.Methodes Twenty-four BALB/C mice were randomly divided into control group,ceftriaxone group and TMC3115 group(n=8).Mice during pregnancy were treated by gavage with normal saline,ceftriaxone(30 mg/d),TMC31115 bacterial suspension(with a bacterial content of 109 CFU/d)daily to delivery,and newborn mice were breast-fed to weaning period(PND21).HE staining was used to observe the depth of colonic villi and crypts.RT-PCR was used to detect the expression of intestinal epithelial tissue development-related proteins(Ki67,Muc2)and tight junction proteins(ZO-1,Claudin-l,Claudin-2,Occludin).The second-generation sequencing was employed to analyze the composition of intestinal flora.Results Compared with control group and ceftriaxone group,the colonic crypt depth and KI67 mRNA expression in the TMC3115 group were significantly higher(P<0.05).In addition,the Occludin mRNA expression in the TMC3115 group was also significantly higher than that of the control group(P<0.05).Firmicutes were the main fecal bacteria in the control and TMC3115 groups.However,Bacteroides was the main fecal bacteria in the Ceftriaxone group.Compared with the Ceftriaxone group,mice in the TMC3115 group had a higher relative abundance of Firmicutes and Lactobacillus(P<0.05)and a lower relative abundance of Clostridium and Enterococcus(P<0.05).Conclusion The treatment of ceftriaxone during pregnancy has no significant effect on the intestinal development of the offspring,but it can significantly damage the structure of the offspring's intestinal flora,which is characterized by the increase of the potential pathogenic bacteria Enterococcus and Clostridium;while the treatment of TMC3115 can significantly increase intestinal Firmicutes and Lactobacillus and promote intestinal tissue development and barrier function.