生物技术通报 ›› 2025, Vol. 41 ›› Issue (2): 77-84.doi: 10.13560/j.cnki.biotech.bull.1985.2024-0783

• 技术与方法 • 上一篇    下一篇

反刍动物乳源microRNA分析技术研究进展

孙同玉1(), 荣飞飞2, 高展2, 马涛1()   

  1. 1.中国农业科学院饲料研究所 农业农村部饲料生物技术重点实验室,北京 100081
    2.山东鲁润畜产品有限公司,德州 250523
  • 收稿日期:2024-08-14 出版日期:2025-02-26 发布日期:2025-02-28
  • 通讯作者: 马涛,男,博士,副研究员,研究方向 :反刍动物营养与调控技术;E-mail: matao@caas.cn
  • 作者简介:孙同玉,男,硕士研究生,研究方向 :反刍动物营养;E-mail: 1612541223@qq.com
  • 基金资助:
    宁夏回族自治区重点研发计划(2023BCF01037);财政部和农业农村部:国家现代农业产业技术体系专项(CARS-38)

Research Advances in the Analytical Technology of Milk-derived microRNA from Ruminant

SUN Tong-yu1(), RONG Fei-fei2, GAO Zhan2, MA Tao1()   

  1. 1.Key Laboratory of Feed Biotechnology of the Ministry of Agriculture and Rural Affairs, Institute of Feed Research, Chinese Academy of Agricultural Sciences, Beijing 100081
    2.Shandong Lurun Animal Products Co. , Ltd, Dezhou 250523
  • Received:2024-08-14 Published:2025-02-26 Online:2025-02-28

摘要:

母乳不仅含有幼龄反刍动物生长必要的营养物质,还含有多种生物活性成分如微小核糖核酸(microRNA)。乳源microRNA存在于胞外囊泡之中,后者是一类脂质双分子层包裹的膜状结构,可以被宿主肠道细胞吸收,对幼龄反刍动物的生长发育起到调控作用,具有一定的抗逆性。分析乳源microRNA组成前通常要先提取胞外囊泡,主要采用差速离心法、密度梯度离心法和沉淀法,每种提取方法都有其优缺点,需要根据样本特点和试验条件选择合适的提取方法,提取到的胞外囊泡还需要经过电子显微镜、流式细胞术或纳米颗粒追踪分析等进行鉴定。乳源microRNA组成分析主要分为测序和生物信息分析两部分,前者需要选择合适的测序平台,后者主要包括数据质控、数据库比对、差异表达microRNA分析和microRNA功能预测等多个步骤,每个步骤都有多种类型分析软件可供选择,需要根据测序数据和分析软件的特点合理搭配分析方法,对测序数据进行精确且有效的信息挖掘。本文主要总结了近年来针对反刍动物乳源microRNA分析技术的研究进展,旨在为合理利用相应的技术研究乳源microRNA的组成和功能,从而有效调控幼龄反刍动物生长发育提供参考。

关键词: 反刍动物, 乳汁, 微小核糖核酸, 小RNA测序

Abstract:

As an essential nutrient, maternal milk also encompasses various bioactive components such as microribonucleic acids (microRNA). microRNA in milk exists in extracellular vesicles (EVs) and can be absorbed into the host's enterocytes, modulating the growth and development of small ruminants, the latter has a membrane structure enclosed by a kind of lipid bilayer with some resistance. Extracellular vesicles derived from milk are often isolated before the analysis of microRNA profiles. Currently, extracellular vesicles are mainly extracted by differential centrifugation, density gradient centrifugation, and precipitation. Each method has specific advantages and disadvantages, an appropriate extraction method should be selected according to sample characteristics and experimental conditions, and extracellular vesicles isolated still need to be identified by electron microscopy, flow cytometry, or nanoparticle tracking analysis. The analysis of microRNA profiles is mainly divided into two parts: Sequencing and data analysis, The former needs to select a suitable sequencing platform according to the requirement, and the latter includes multiple steps such as data quality control, alignment with the database, differential expression microRNA analysis and microRNA function prediction. For each step, there are various types of analysis software to choose from, analysis methods should be reasonably combined according to the characteristics of sequencing data and software to carry out more accurate and effective information mining on sequencing data. this paper reviewed the research advances in the ruminants' microRNA, aiming to use corresponding technology to research the profiles and function of microRNA derived from milk, providing references for modulating the growth and development of small ruminants.

Key words: ruminant, milk, microRNA, small RNA sequencing