孕前和孕期高脂饲料喂养对新生大鼠肠道菌群的影响  

Effects of high-fat diet in pre-pregnancy and pregnancy on gut microbiota of neonatal rats

作　　者：胡卓 杨倩 魏宇辰 蔡夏夏 党秦豫 朱砚荻 张亚迪 陈朝阳 余焕玲 HU Zhuo;YANG Qian;WEI Yu-chen;CAI Xia-xia;DANG Qin-yu;ZHU Yan-di;ZHANG Ya-di;CHEN Zhao-yang;YU Huan-ling(Department of Nutrition and Food Hygiene,College of Public Health,Capital Medical University,Beijing 100069,China)

机构地区：[1]首都医科大学公共卫生学院营养与食品卫生学系,北京100069

出　　处：《现代预防医学》2024年第4期697-704,共8页Modern Preventive Medicine

基　　金：北京市教育委员会科技/社科计划项目(KZ202210025039)。

摘　　要：目的 探讨孕前和孕期高脂饲料喂养对新生大鼠肠道菌群的影响。方法 12只3周龄无特定病原体(Specific Pathogen Free, SPF)级雌性Sprague-dawley(SD)大鼠,适应性喂养一周后,按体重随机分为对照组与高脂组,孕前分别用基础饲料与高脂饲料喂养至12周龄,与同周龄SPF级雄性SD大鼠合笼,次日观察到阴道栓或阴道涂片中发现精子后,判定受孕。孕鼠单笼饲养,孕期的饲料与孕前相同。收集出生后第0.5天(postnatal day 0.5,PD 0.5)仔鼠,称量体重,采集肠道组织并称量质量,计算肠道系数;进行回肠和结肠组织病理和相关基因检测;提取肠道内容物的细菌总DNA,通过16S rRNA测序分析肠道菌群。采用t检验或Mann-Whitney U检验分析数据。结果 与对照组相比,高脂组PD 0.5仔鼠体重、肠道质量、肠道系数无显著改变(t=-0.682,P=0.499;t=0.269,P=0.790;t=0.674,P=0.506);肠道病理形态无改变;结肠闭合蛋白-1(Claudin-1)、溶菌酶(Lysozyme)、再生胰岛衍生蛋白3伽马(regenerating islet derived protein 3 gamma, Reg3γ)和α-防御素5(Alpha-defensin 5)的基因表达水平显著增加(t=-3.136,P=0.011;t=-3.303,P=0.019;t=-3.093,P=0.011;t=-3.575,P=0.005);肠道菌群的α多样性和β多样性无显著改变(t=-1.407,P=0.190;t=-1.410,P=0.189;t=-2.017,P=0.071;U=6.000,P=0.055;F=1.030,P=0.141);肠道瘤胃球菌属、考拉杆菌属、Lachnospiracea_incertae_sedis、韦荣氏球菌属、黄杆菌属和Bacteroides_sp._SLC1-38的丰度显著增加(U=3.500,P=0.013;U=7.000,P=0.050;U=6.000,P=0.022;U=6.000,P=0.022;U=7.000,P=0.050;U=3.000,P=0.007);线性判别分析效应量(Linear discriminant analysis Effect Size, LEfSe)分析表明高脂组中对差异影响贡献较大的物种有埃希氏菌属/志贺氏菌属、瘤胃球菌属、考拉杆菌属、Lachnospiracea_incertae_sedis和Bacteroides_SP._SLC1-38。结论 孕前和孕期高脂饲料喂养导致PD 0.5仔鼠肠道代谢相关菌和致病菌的丰度增加。Objective To investigate the effects of high-fat diet in pre-pregnancy and pregnancy on gut microbiota of neonatal rats.Methods Twelve 3-week-old Specific Pathogen Free(SPF)female Sprague-dawley(SD)rats after one week of adaptive feeding were randomly divided into control group and high-fat diet group according to body weight.The rats were fed basal diet or high-fat diet until 12-week-old in pre-pregnancy,respectively,and then mated with SPF male SD rats of the same age.Pregnancy was determined the next day when vaginal plugs were observed or sperms were found by vaginal smears.Pregnant rats were kept in single cages and the diet in pregnancy was the same as pre-pregnancy.The postnatal day 0.5(PD 0.5)neonatal rats were collected and the body weights were measured.Intestine of PD 0.5 neonatal rats were collected,intestinal weights were measured and intestinal coefficients were calculated.Histopathology and related genes detection of ileum and colon were performed.Total bacterial DNA of intestinal contents was extracted and the gut microbiota was analyzed by 16S rRNA sequencing.Data was analyzed by t test or Mann-Whitney U test.ResultsCompared with control group,there were no significant difference in the body weight,intestinal weight and intestinal coefficient in PD O.5 neonatal rats of high-fat diet group(t=-0.682,P=0.499;t=0.269,P=0.790;t=0.674,P=0.506);there were no difference in intestinal histopathological changes;the gene expression level of Claudin-1,Lysozyme,Reg3(regenerating islet derived protein 3 gamma)and Alpha-defensin 5 of colon in PD 0.5 neonatal rats of high-fat diet group were increased significantly(t=-3.136,P=0.011;t=-3.303,P=0.019;t=-3.093,P=0.011;t=-3.575,P=0.005);there were no significant difference inαdiversity andβdiversity of gut microbiota(t=-1.407,P=0.190;t=-1.410,P=0.189;t=-2.017,P=0.071;U=6.000,P=0.055;F=1.030,P=0.141);the abundance of Ruminococcus,Phascolarctobacterium,Lachnospiracea_incertae_sedis,Veillonella and Flavobacterium and Bacteroides_sp._SLC1-38 increased significantly(U=

关 键 词：高脂饮食 子代 肠道菌群 16S rRNA 

分 类 号：R117[医药卫生—公共卫生与预防医学]