孕前和孕哺期小鼠高脂饮食对子代肠道菌群的影响  

Effects of high-fat diet in pre-pregnancy and pregnancy-and-lactation periods on gut microbiota of offspring mice

作　　者：许锐 洪芳兰 陈晓伟 吴蕾蕾 沈孝兵 XU Rui;HONG Fanglan;CHEN Xiaowei;WU Leilei;SHEN Xiaobing(Key Laboratory of Environmental Medical Engineering of Ministry of Education/School of Public Health,Southeast University,Nanjing,Jiangsu 210009,China)

机构地区：[1]东南大学,公共卫生学院/环境医学工程教育部重点实验室,江苏南京210009

出　　处：《环境与职业医学》2022年第8期849-855,共7页Journal of Environmental and Occupational Medicine

摘　　要：[背景]近年来研究发现肠道菌群在各种慢性疾病的发生发展中起着重要的作用,饮食是影响肠道菌群的重要因素。然而,关于不同时期母体高脂饮食对子代肠道微生物影响的研究仍然有限。[目的]探讨母鼠孕前期和孕哺期高脂饮食对子代肠道微生物群的影响。[方法]根据孕前期和孕哺期给予的不同饮食(高脂饮食,HFD;对照饮食,CD),将C57BL/6J雌性小鼠分为四组,分别是孕前对照饮食—孕哺期对照饮食组(CD-CD组)、孕前对照饮食—孕哺期高脂饮食组(CD-HFD组)、孕前高脂饮食—孕哺期对照饮食组(HFD-CD组)、孕前高脂饮食—孕哺期高脂饮食组(HFD-HFD组)。母鼠在喂养6周后与雄性小鼠同笼受孕,将成功受孕的母鼠继续分为两组,给予不同的饮食。子鼠出生后直接由母鼠母乳喂养,每只母鼠仅哺乳1只子代小鼠,最后每组子鼠数量为6只,雌雄各半。监测记录后代小鼠体重,比较各组体重增长差异。哺乳期结束后收集子鼠新鲜粪便,提取粪便中细菌总DNA,根据细菌16S rDNA(V3+V4)序列设计特异性引物进行扩增,接着基于Illumina HiSeq 2500平台进行测序,利用QIMME、USEARCH、R软件对测序数据进行注释及操作分类单元(OTU)分析并绘制物种丰度柱状图。α多样性分析中ACE和Chao1指数衡量菌群的物种丰富度,Shannon和Simpson指数综合考量丰富度与均匀度;β多样性分析中主坐标分析(PCoA)和相似性分析(Anosim)比较各组间菌群组成差异,线性判别分析(LEfSe)用于寻找对各组间差异影响较大的菌群。[结果]子鼠体重变化结果显示孕哺期的饮食对后代的影响更大,整个实验周期内HFD-CD组体重最低。OTU分析表明孕哺期高脂饮食降低了子鼠的OTU数量,α多样性分析显示孕哺期高脂饮食降低了子代肠道菌群丰富度(ACE,P<0.05;Chao1,P<0.05),而孕前高脂饮食的子鼠各α多样性指数无明显差异。不同时期的高脂饮食还导致了子鼠优势[Background]In recent years gut microbiota has been found to play an important role in the occurrence and development of various chronic diseases,and diet is an important factor influencing gut microbiota.However,the effects of maternal high-fat diet in pre-pregnancy and pregnancy-and-lactation periods on offspring gut microbiota are still unclear.[Objective]To investigate the effects of maternal high-fat diet in pre-pregnancy and pregnancy-and-lactation periods on gut microbiota of offspring mice.[Methods]C57BL/6J female mice were divided into four groups according to the diet patterns(high-fat diet,HFD;control diet,CD)given before and after conception,namely the pre-pregnancy control diet and post-pregnancy control diet group(CD-CD group),the pre-pregnancy control diet and post-pregnancy high-fat diet group(CD-HFD group),the pre-pregnancy high-fat diet and post-pregnancy control diet group(HFD-CD group),and the pre-pregnancy high-fact diet and post-pregnancy high-fat diet group(HFD-HFD group).Female mice were conceived in the same cage with male mice after 6 weeks of feeding,and the successfully conceived females continued to be randomly divided into two groups receiving either high-fat or control diet,and when the offspring mice were born,they were breastfed directly by the mothers,with each mother nursing only one offspring mouse.The number of offspring mice in each group was 6,with half males and half females.The body weight of offspring mice were recorded and body weight gain was compared between the four groups.After the lactational period,fresh feces of the offspring were collected,and the fecal DNA was extracted.Specific primers were designed according to the bacterial 16S rDNA(V3+V4)sequence and then the sequencing was performed using the Illumina HiSeq 2500 platform.Species annotation and operational taxonomic unit(OTU)analysis of sequencing data were conducted using QIMME,USEARCH and R software.In alpha diversity analysis,ACE and Chao1 indices were used to evaluate species richness,Shannon and Simpson

关 键 词：孕前期 孕哺期 小鼠 高脂饮食 肠道菌群 16SrRNA 多样性分析 

分 类 号：R117[医药卫生—公共卫生与预防医学]