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Mammary tissue proteomics in a pig model indicates that dietary valine supplementation increases milk fat content via increased de novo synthesis of fatty acid

ABSTRACT: Abstract Milk fat is a major source of energy that determines the growth of neonates. Recently, studies have shown that valine is closely related to lipid metabolism. Therefore, this study was designed to investigate the effects of dietary valine supplementation on milk fat synthesis using a pig model. Thirty gilts were allotted to low (LV, total valine:lysine = 0.63:1), medium (MV, total valine:lysine = 0.73:1), and high (HV, total valine:lysine = 0.93:1) valine feeding levels from Day 75 of gestation till farrowing. The results demonstrated that the concentration of milk fat at Days 1, 3, and 7 of lactation in the HV group was higher than that in the MV and LV groups. The HV group had an increased (p < .05) proportion of total saturated and monounsaturated fatty acids than the other groups. Examination of mammary tissue proteomics in the HV and LV groups revealed 121 differentially expressed proteins (68 upregulated and 53 downregulated in the HV group). The upregulated proteins in the HV group were relevant to some crucial pathways related to milk fat synthesis, including fatty acid biosynthesis and metabolism, the AMPK signaling pathway, and oxidative phosphorylation. Furthermore, the key proteins involved in fatty acid synthesis (ACACA and FASN) were identified, and their expression levels were verified (p < .05) using Western blotting. Our findings revealed that dietary valine supplementation improves milk fat synthesis by modulating the expression of fatty acid synthesis–related proteins in mammary tissues. Dietary valine supplementation increases the content of milk fat in the colostrum Dietary valine supplementation increases the proportion of fatty acids in colostrum. Valine can activate the signaling pathway related to fatty acids synthesis. Valine promote fatty acids synthesis may via the upregulation of proteins ACACA and FASN

SUBMITTER: Che L

PROVIDER: S-EPMC8565212 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:To investigate the role of glucose in regulating milk fatty acid synthesis, 6 lactating Guanzhong dairy goats were infused with 0, 60, or 100 g/d glucose via the external pubic artery in a 3 × 3 repeated Latin square experiment. A concomitant in vitro experiment was conducted to investigate possible mechanisms whereby glucose regulates milk fatty acid synthesis. RNA sequencing was used for cellular transcriptome analysis. Drugs, MK-2206, rapamycin, and dorsomorphin were used to block cellular mammalian AMP-activated protein kinase (AMPK), AKT serine/threonine kinase 1, and mechanistic target of rapamycin kinase signaling pathways, respectively. Carbohydrate response element binding protein (ChREBP) was knockdown and overexpressed to investigate its role in regulating milk fatty acid synthesis in mammary epithelial cells. Glucose infusion linearly elevated the concentration of C8:0 (P = 0.039) and C10:0 (P = 0.041) in milk fat while it linearly decreased (P = 0.049) that of C16:0. This result was in agreement with the upregulation of genes related to de novo synthesis of fatty acids and lipid droplet formation, including adipose differentiation-related protein, butyrophilin subfamily 1 member A1, fatty acid synthase (FASN) and ChREBP. Their expression increased (P < 0.05) linearly in the lactating goat mammary gland. In vitro, glucose linearly stimulated the expression of genes related to de novo synthesis of fatty acids and cellular triacylglycerol in cultured mammary epithelial cells. RNA sequencing and inhibition studies revealed that glucose induced transcriptomic changes increasing lipogenic pathways, with AMPK responding to glucose by controlling ChREBP and FASN. Knockdown and overexpression of ChREBP highlighted its essential role in lipogenesis. The knockdown and overexpression of ChREBP protein also revealed an essential role in regulating the de novo synthesis of fatty acids. Collectively, our data highlight that glucose supplementation promotes de novo fatty acid synthesis via the AMPK-ChREBP axis, hence increasing milk fat yield in the goat mammary gland. Results from the current study provide possible strategies to manipulate the fatty acid composition as well as improve ruminant milk quality.

Project description:Background:Mammary gland development during late gestation in gilts is a major factor that alters the composition of colostrum and growth performance of piglets. Plasma valine is taken up and metabolized extensively by the mammary gland; however, the effects of valine on mammary gland development during late gestation are still unclear. Thirty primiparous gilts were divided into three treatment groups (n =?10) and received one of the three diets starting on day 75 of gestation until the day of farrowing. The total dietary valine to lysine ratio of the three diets was 0.63 (LV), 0.73 (MV), and 0.93 (HV), respectively. Results:Dietary valine supplementation during late gestation did not affect (P >?0.05) the litter size and weight at farrowing; however, the piglet weight and average daily gain at weaning were linearly increased (P <?0.05) as the dietary valine increased. The highest piglet weight at weaning was observed when the gilts were provided the HV diet. Dietary valine supplementation linearly elevated (P <?0.05) protein, fat and solids-not-fat and some free amino acids content in colostrum. The concentration of prolactin in plasma of gilts was linearly increased in response to valine supplementation at days 1 and 10 of lactation (P <?0.05). Furthermore, with increasing dietary valine allowance, a linear increase (P <?0.05) was observed in the area of the lumen of alveolus and the content of DNA, RNA, and total protein in the mammary tissues at day 1 of lactation. Moreover, the protein expression of cyclin D1, p-mTOR, p-S6, and p-4EBP1 was also linearly increased (P <?0.05) in the mammary tissue at day 1 of lactation. However, no difference (P >?0.05) was observed in the indices related to mammary development and the mTOR signaling pathway at day 21 of lactation. Conclusion:The results revealed that increasing the total dietary valine to lysine ratio to 0.93 during late gestation significantly enhances the piglet weight and average daily gain at weaning probably due to improved development of mammary gland.

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Mammary tissue proteomics in a pig model indicates that dietary valine supplementation increases milk fat content via increased de novo synthesis of fatty acid

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