应用Illumina NovaSeq测序技术比较3种杂粮对大鼠肠道菌群的影响

Effects of Three Kinds of Coarse Cereals on Gut Microbiota of Rats Explored by Illumina NovaSeq Sequencing Technology

为探究燕麦、荞麦和小米对健康大鼠肠道结构、肠道菌群及肠道中短链脂肪酸(short-chain fatty acids,SCFAs)含量的影响,本实验将48只SPF级雄性SD大鼠随机分为4组(空白对照组(饲喂标准维持饲料)、燕麦组(饲喂含22%(质量分数,下同)燕麦的饲料)、荞麦组(饲喂含22%荞麦的饲料)和小米组(饲喂含22%小米的饲料)),每周测定大鼠体质量,12周后处死大鼠,取肝脏、结肠组织及结肠内容物,对大鼠肝脏和结肠组织进行病理学检测;应用Illumina NovaSeq高通量测序技术检测大鼠肠道菌群变化;利用液相色谱-质谱联用仪检测大鼠结肠内容物SCFAs含量。结果表明,燕麦可增加大鼠肠道菌群多样性,提高大鼠结肠乳杆菌属(Lactobacillus)和阿克曼氏菌属(Akkermansia)丰度;荞麦和小米可增加厚壁菌门(Firmicutes)相对丰度,降低疣微菌门(Verrucomicrobia)和拟杆菌门(Bacteroidetes)相对丰度;燕麦、荞麦和小米均可降低拟杆菌属(Bacteroides)丰度;燕麦和小米可显著提高大鼠结肠内乙酸和总SCFAs含量(P<0.05),小米可显著提高丙酸和异丁酸含量(P<0.05)。综上,燕麦对大鼠肠道菌群具有一定的改善作用,荞麦和小米对肠道菌群的影响相似度较高,相关研究结果可为谷物功能食品的开发提供科学依据。

To investigate the effect of dietary supplementation with oats,tartary buckwheat and foxtail millet on the intestinal histology,microbiota and short-chain fatty acids(SCFAs)in normal rats,48 male SD rats were randomly and equally divided into four groups:control(fed a standard maintenance diet),oat(fed the maintenance diet containing 22%oats),tartary buckwheat(fed the maintenance diet containing 22%tartary buckwheat)and foxtail millet groups(fed the diet containing 22%foxtail millet).Body mass was recorded weekly.After 12 weeks of feeding,the rats were sacrificed to collect liver tissue,colon tissue and colonic contents.Histopathological characteristics of liver and colon tissues were observed.The intestinal microbiota was analyzed by Illumina NovaSeq high-throughput sequencing technology.SCFAs in colonic contents were determined by liquid chromatography-mass spectrometry.Results indicated that administration of oats increased gut microbiota diversity and showed higher relative abundance of Lactobacillus and Akkermansia than the control group.Tartary buckwheat and foxtail millet increased the relative abundance of Firmicutes,but decreased the abundance of Verrucomicrobia and Bacteroidetes in the gut of normal rats.Oats,tartary buckwheat and foxtail millet decreased the relative abundance of Bacteroides.Oats and foxtail millet could significantly increase the concentrations of acetic acid and total SCFAs(P<0.05),and foxtail millet could significantly increase the concentrations of propionic acid and isobutyric acid in colonic contents(P<0.05).In conclusion,oat supplementation has regulatory effects on the gut microbiota in rats,and tartary buckwheat and foxtail millet have similar effects on it.This study provides a scientific basis for the development of cereal-based functional foods.