Journal of Integrative Agriculture ›› 2022, Vol. 21 ›› Issue (10): 3004-3016.DOI: 10.1016/j.jia.2022.07.039

所属专题： 动物科学合辑Animal Science

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JIA-2021-1115 MiR-140通过靶向牛乳腺上皮细胞中的转化生长因子α(TGFA)下调脂肪酸的合成

  

  • 收稿日期:2021-06-28 接受日期:2022-05-07 出版日期:2022-10-01 发布日期:2022-05-07

MiR-140 downregulates fatty acid synthesis by targeting transforming growth factor alpha (TGFA) in bovine mammary epithelial cells

CHU Shuang-feng1, 2, ZHAO Tian-qi1, 2, Abdelaziz Adam Idriss ARBAB1, 2, YANG Yi2, 3, CHEN Zhi1, 2, YANG Zhang-ping1, 2   

  1. 1College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, P.R.China

    2Joint International Research Laboratory of Agriculture & Agri-Product Safety, Yangzhou University, Yangzhou 225009, P.R.China

    3Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, P.R.China

  • Received:2021-06-28 Accepted:2022-05-07 Online:2022-10-01 Published:2022-05-07
  • About author: CHU Shuang-feng, Tel: +86-15861327537, E-mail: DX120200142@yzu.edu.cn; Correspondence YANG Zhang-ping, Tel: +86-514-87979307, E-mail: yzp@yzu.edu.cn; CHEN Zhi, E-mail: zhichen@yzu.edu.cn. *These authors contributed equally to this study.
  • Supported by:

    This research was supported by the National Natural Science Foundation of China (31802035, 31872324 and 31601915).

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摘要：

脂肪是维持生命不可缺少的营养物质和基本代谢产物，牛奶中富含脂肪酸，包括多种饱和脂肪酸和不饱和脂肪酸。miRNA和mRNA在乳腺组织乳脂代谢的调控中起重要作用，有研究表明，脂质代谢具有复杂的转录调控作用，但人们对通过miRNA-mRNA相互作用调控乳脂合成的机制知之甚少。本研究以泌乳后期（分娩后270d和315d）的牛乳腺组织进行转录组测序，鉴定调控乳脂代谢的关键基因，共筛选出1207个差异共表达基因，其中包括828个上调基因和379个下调基因。选择转化生长因子-α (TGFA) 基因为本实验的靶向目的基因，采用荧光素酶报告基因、Western blotting和qRT-PCR检测进行进一步的功能研究。结果表明了miR-140是TGFA的上游调控因子，miR-140能抑制(P < 0.01)牛乳腺上皮细胞 (BMECs) 和甘油三酯(TAGs)产生；相反，TGFA促进(P < 0.01)不饱和脂肪酸和TAG生成。拯救实验进一步表明了miR-140/TGFA调控机制。综上所述，这些结果表明了miR-140/TGFA通路可以抑制(P < 0.01)奶牛乳腺上皮细胞中的乳脂代谢的进行，通过遗传手段改善牛奶品质。本研究的创新点主要表现为：目前，国内外的研究大部分集中于四个泌乳阶段的研究，而很少有人专注于两个泌乳阶段的研究，而且大家的研究方向都倾向于泌乳初期，盛期和中期的研究，很少有人把注意力放在泌乳后期的两个阶段，以至于奶牛泌乳后期乳脂代谢的大数据都很少，不能使得牧场工作人员对奶牛饲养工作有更全面的了解，而本研究集中于泌乳后期的两个阶段的研究，有效的帮助牧场工作人员对泌乳后期奶牛体内脂质代谢知识的了解，也弥补了国内外在此阶段研究缺失的遗憾。

Abstract:

Fat is an indispensable nutrient and basic metabolite for sustaining life, and milk is particularly rich in fatty acids, including a variety of saturated and unsaturated fatty acids.  MicroRNA (miRNA) and mRNA play an important role in the regulation of milk fat metabolism in mammary gland tissue.  It has been shown that lipid metabolism has a complex transcriptional regulation, but the mechanism by which milk fat synthesis is regulated through miRNA–mRNA interactions is poorly understood.  In this study, we performed transcriptome sequencing with bovine mammary gland tissue in the late lactation (270 and 315 days after parturition) to identify the key gene that regulating milk fat metabolism.  A total of 1 207 differentially coexpressed genes were selected, 828 upregulated genes and 379 downregulated genes were identified.  The transforming growth factor alpha (TGFA) gene was selected as the target gene, and luciferase reporter assay, Western blotting and qRT-PCR were used for further study.  The results demonstrated that miR-140 was an upstream regulator of TGFA, and miR-140 could inhibit (P<0.01) unsaturated fatty acid and triglyceride (TAGs) production in bovine mammary epithelial cells (BMECs).  In contrast, TGFA promoted (P<0.01) unsaturated fatty acid and TAG production.  Rescue experiments further indicated the miR-140/TGFA regulatory mechanism.  Taken together, these results suggest that the miR-140/TGFA pathway can inhibit (P<0.01) milk fat metabolism and improve milk quality by genetic means.

Key words: bovine mammary epithelial cells ,  TGFA , transcriptome sequencing , miR-140 , fatty acid metabolism