生物技术通报 ›› 2022, Vol. 38 ›› Issue (5): 159-168.doi: 10.13560/j.cnki.biotech.bull.1985.2021-1071

• 研究报告 • 上一篇    下一篇

miR-665在奶牛乳腺上皮细胞炎症中的表达及功能分析

李宇航1,2(), 王兴平1,2(), 杨箭1,2, 罗仍卓么1,2, 任倩倩1,2, 魏大为1,2, 马云1,2   

  1. 1.宁夏大学农学院,银川 750021
    2.宁夏回族自治区反刍动物分子细胞育种重点实验室,银川 750021
  • 收稿日期:2021-08-20 出版日期:2022-05-26 发布日期:2022-06-10
  • 作者简介:李宇航,女,硕士,研究方向:动物分子遗传育种;E-mail: 1429692080@qq.com
  • 基金资助:
    国家自然科学基金项目(31960652);国家自然科学基金项目(32060749);宁夏回族自治区重点研发计划(引才专项)项目(2019BEB04002);宁夏高等学校科学研究项目(NGY2020007);宁夏大学引进人才科研启动项目(030900001926)

Expression and Functional Analysis of miR-665 in Bovine Mammary Epithelial Cell Inflammation

LI Yu-hang1,2(), WANG Xing-ping1,2(), YANG Jian1,2, LUORENG Zhuo-ma1,2, REN Qian-qian1,2, WEI Da-wei1,2, MA Yun1,2   

  1. 1. School of Agriculture,Ningxia University,Yinchuan 750021
    2. Key Laboratory of Ruminant Molecular Cell Breeding,Ningxia Hui Autonomous Region,Yinchuan 750021
  • Received:2021-08-20 Published:2022-05-26 Online:2022-06-10

摘要:

为探究miR-665在奶牛乳腺上皮细胞炎症中的表达及功能,利用脂多糖(LPS)诱导奶牛乳腺上皮细胞产生炎症反应,在LPS诱导0、3、6和12 h采用qPCR技术检测了miR-665及其潜在靶mRNA的表达水平,并采用生物信息学方法进行了miR-665保守性分析、靶基因预测及其KEGG和GO功能富集分析。结果表明,miR-665在牛、人和小鼠等物种间高度保守;与0 h相比,miR-665在LPS诱导乳腺上皮细胞产生炎症的3、6和12 h的表达量均显著上调,且与促进炎症相关的潜在靶基因(MAPK14MAP3K2SMAD2SP1)的表达量呈相反趋势;miR-665的靶基因预测及其KEGG和GO功能富集结果显示,miR-665可能通过参与MAPK、Th17细胞分化、肿瘤坏死因子等信号通路发挥作用。推测miR-665可能在LPS诱导的乳腺上皮细胞炎症中具有抑制作用。

关键词: 奶牛, miR-665, 乳腺上皮细胞, 乳腺炎, 生物信息学分析

Abstract:

In order to explore the expression and function of miR-665 in bovine mammary epithelial cell inflammation,lipopolysaccharide(LPS)was used to induce bovine mammary epithelial cell inflammation,qPCR technology was applied to detect the expression levels of miR-665 and its potential target mRNA at 0,3,6 and 12 h of LPS-induced. Bioinformatics method was adapted to conduct the conserved miR-665,target gene prediction,KEGG and GO function enrichment analysis. The results showed that miR-665 was highly conserved in many species,such as Bos taurus,Homo sapiens and Mus musculus. Compared with 0 h,the expression levels of miR-665 were significantly up-regulated at 3,6 and 12 h of LPS-induced inflammation in the mammary epithelial cells,while the expression levels of MAPK14,MAP3K2,SMAD2,and SP1 genes related to inflammation promotion were in an opposite trend. The predicted target genes of miR-665 and the enrichment results of KEGG and GO functions showed that miR-665 may play its role by participating in MAPK and Th17 cell differentiation,tumor necrosis factor and other signaling pathways. It is speculated that miR-665 may have an inhibitory effect on the LPS-induced inflammation of mammary epithelial cells.

Key words: dairy cow, miR-665, mammary epithelial cell, mastitis, bioinformatics analysis