食品科学 ›› 2020, Vol. 41 ›› Issue (6): 155-162.doi: 10.7506/spkx1002-6630-20181227-317

• 生物工程 • 上一篇    下一篇

基于16S rRNA技术分析α-乳白蛋白对大鼠肠道菌群的影响

李梦寒,王志勇,盛雪,崔东影,席恩泽,汤梦琪,张惠恺,许晓曦,马春丽   

  1. (1.东北农业大学食品学院,黑龙江 哈尔滨 150030;2.黑龙江农业工程职业学院,黑龙江 哈尔滨 150088)
  • 出版日期:2020-03-25 发布日期:2020-03-23
  • 基金资助:
    “十二五”国家科技支撑计划项目(2013BAD18B00)

Effect of α-Lactalbumin on Intestinal Flora in Rats Analyzed by 16S rRNA High-Throughput Sequencing

LI Menghan, WANG Zhiyong, SHENG Xue, CUI Dongying, XI Enze, TANG Mengqi, ZHANG Huikai, XU Xiaoxi, MA Chunli   

  1. (1. College of Food Science, Northeast Agricultural University, Harbin 150030, China;2. Heilongjiang Agricultural Engineering Vocational College, Harbin 150088, China)
  • Online:2020-03-25 Published:2020-03-23

摘要: 探究不同添加量α-乳白蛋白(α-lactalbumin,α-LA)对肠道菌群多样性及其结构的影响,研究婴儿配方奶粉中α-LA科学添加量十分重要。本实验将4 组SD大鼠分别灌喂等剂量的生理盐水(对照组)、高添加量α-LA(200 mg/kg)、中添加量α-LA(100 mg/kg)、低添加量α-LA(20 mg/kg),在灌喂第14、28天时无菌取结肠粪便,扩增肠道菌群16S rRNA基因的V3-V4区,通过高通量测序分析α多样性、β多样性及基于门、属水平的物种组成。结果发现,高添加量蛋白组、中添加量蛋白组提高了肠道菌群的丰富度、均匀度,更好地调整了菌群结构。在门水平上,24 份样品的肠道菌群均以厚壁菌门和拟杆菌门为主,各添加量蛋白组对于门水平的调节与母乳相似。在属水平上,高添加量蛋白组会降低乳杆菌、布鲁氏菌、阿克曼氏菌的相对丰度,而中、低蛋白组有一定的促进作用;各添加量组均能使普氏菌属成员、拟杆菌的相对丰度有不同程度的降低,作用效果随添加量降低而提高;各添加量组均能提高木质真菌相对丰度并维持瘤胃球菌科、毛螺旋菌科成员、梭菌科下的未知属相对丰度恒定。中添加量蛋白在提高菌群多样性的同时,提高或维持了肠道益生菌的相对丰度,同时降低了肠道有害菌的丰度,为最佳实验添加量。本实验结果可为婴儿喂养实验提供参考,为婴儿配方奶粉中α-LA科学的添加量提供理论支持。

关键词: α-乳白蛋白, 16S?rRNA, 婴儿, 肠道菌群, SD大鼠

Abstract: Alpha-lactalbumin (α-LA) is an important immune protein in breast milk. In addition to providing various nutrients such as essential amino acids for infants, α-LA may also regulate intestinal flora. Infant formula contains a lower concentration of α-LA relative to breast milk. Therefore, it is important to explore the effects of different concentrations of α-LA on the diversity and structure of intestinal flora and to determine the appropriate amount of α-LA science added in infant formula. In this experiment, 4 groups of SD rats were orally administered respectively with an equal volume of normal saline (control group), α-LA at high, medium and low concentrations (200, 100 and 20 mg/kg), and colonic feces were aseptically taken after 14 and 28 days of administration. The V3-V4 region of the 16S rRNA gene of the intestinal flora was amplified. Alpha diversity, Beta diversity, and intestinal flora composition at the phylum and genus levels were analyzed by high-throughput sequencing. The results showed that α-LA at high and medium doses improved the species richness, diversity and structure of intestinal microbiota, and the effect was similar to that of breast milk. At the genus level, the high concentration of α-LA reduced the relative abundance of Lactobacillus, Blautia and Akkermansia, while the opposite effect was observed at medium and low concentrations. α-LA at all concentrations decreased the relative abundance of Prevotella and Bacteroides, and the effect was more pronounced with the decrease in its concentration. Moreover, α-LA at all concentrations increased the relative abundance of Eubacterium xylanophilum and maintained the relative abundance of Ruminococcaceae, Clostridium sensu stricto_1 and unknown Lachnospiraceae genera at a constant level. The medium concentration of α-LA enhanced intestinal microbial diversity, and increased or maintained the relative abundance of intestinal probiotics while reducing the abundance of intestinal harmful bacteria, suggesting α-LA at this concentration was optimal to regulate intestinal microbiota. The results of this study provide experimental data for improving infant feeding, and ultimately provide theoretical support for the supplementation of α-LA in infant formula.

Key words: α-lactalbumin, 16S rRNA, infant, intestinal flora, SD Rat

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