Université Clermont Auvergne,INRA, VetAgro Sup, UMR Herbivores, F-63122 Saint-Genès-Champanelle, France.
Department of Pathology and Veterinary Diagnostics, Faculty of Veterinary Medicine, Warsaw University of Life Sciences,02-776 Warsaw, Poland.
Int J Mol Sci. 2019 Mar 6;20(5):1156. doi: 10.3390/ijms20051156.
The objective is to study the effects of nutrient restrictions, which induce a metabolic imbalance on the inflammatory response of the mammary gland in early lactation cows. The aim is to decipher the molecular mechanisms involved, by comparing a control, with a restriction group, a transcriptome and proteome, after an intra-mammary lipopolysaccharide challenge. Multi-parous cows were either allowed intake of a lactation diet ( = 8), or a ration containing low nutrient density ( = 8; 48% barley straw and dry matter basis) for four days starting at 24 ± 3 days in milk. Three days after the initiation of their treatments, one healthy rear mammary quarter of 12 lactating cows was challenged with 50 µg of lipopolysaccharide (LPS). Transcriptomic and proteomic analyses were performed on mammary biopsies obtained 24 h after the LPS challenge, using bovine 44K microarrays, and nano-LC-MS/MS, respectively. Restriction-induced deficits in energy, led to a marked negative energy balance (41 versus 97 ± 15% of Net Energy for Lactation (NEL) requirements) and metabolic imbalance. A microarray analyses identified 25 differentially expressed genes in response to restriction, suggesting that restriction had modified mammary metabolism, specifically β-oxidation process. Proteomic analyses identified 53 differentially expressed proteins, which suggests that the modification of protein synthesis from mRNA splicing to folding. Under-nutrition influenced mammary gland expression of the genes involved in metabolism, thereby increasing β-oxidation and altering protein synthesis, which may affect the response to inflammation.
本研究旨在探讨营养限制(导致代谢失衡)对泌乳早期奶牛乳腺炎症反应的影响。为此,我们比较了对照组和限制组,在经乳腺内脂多糖(LPS)刺激后,进行转录组和蛋白质组分析,以阐明其中涉及的分子机制。选取经产奶牛,在泌乳期,自由采食泌乳日粮( = 8 头)或低营养密度日粮( = 8 头,干物质基础中 48%为大麦秸秆),限制组奶牛从泌乳 24 ± 3 天开始,连续 4 天限制采食。处理 3 天后,对 12 头泌乳奶牛的 12 个健康后乳区中的每一个进行 LPS(50 µg)挑战。LPS 刺激后 24 小时,通过牛 44K 微阵列和纳升液相色谱-串联质谱(nano-LC-MS/MS),分别对乳腺活检组织进行转录组和蛋白质组分析。能量限制导致能量严重负平衡(限制组奶牛为 41%,而泌乳净能需求为 97 ± 15%)和代谢失衡。微阵列分析发现,25 个基因在限制条件下差异表达,表明限制改变了乳腺代谢,特别是β-氧化过程。蛋白质组分析鉴定到 53 个差异表达蛋白,表明从 mRNA 剪接到折叠,蛋白质合成过程发生了改变。营养不足影响了乳腺代谢相关基因的表达,从而增加了β-氧化和改变了蛋白质合成,这可能会影响炎症反应。