Dataset Information

Plasma proteomics reveals crosstalk between lipid metabolism and immunity in dairy cows receiving essential fatty acids and conjugated linoleic acid.

ABSTRACT: Essential fatty acids (EFA) and conjugated linoleic acids (CLA) are unsaturated fatty acids with immune-modulatory effects, yet their synergistic effect is poorly understood in dairy cows. This study aimed at identifying differentially abundant proteins (DAP) and their associated pathways in dairy cows supplied with a combination of EFA and CLA during the transition from antepartum (AP) to early postpartum (PP). Sixteen Holstein cows were abomasally infused with coconut oil as a control (CTRL) or a mixture of EFA (linseed + safflower oil) and CLA (Lutalin, BASF) (EFA + CLA) from - 63 to + 63 days relative to parturition. Label-free quantitative proteomics was performed on plasma samples collected at days - 21, + 1, + 28, and + 63. During the transition time, DAP, consisting of a cluster of apolipoproteins (APO), including APOE, APOH, and APOB, along with a cluster of immune-related proteins, were related to complement and coagulation cascades, inflammatory response, and cholesterol metabolism. In response to EFA + CLA, specific APO comprising APOC3, APOA1, APOA4, and APOC4 were increased in a time-dependent manner; they were linked to triglyceride-enriched lipoprotein metabolisms and immune function. Altogether, these results provide new insights into metabolic and immune adaptation and crosstalk between them in transition dairy cows divergent in EFA + CLA status.

SUBMITTER: Veshkini A

PROVIDER: S-EPMC8983735 | biostudies-literature | 2022 Apr

REPOSITORIES: biostudies-literature

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Plasma proteomics reveals crosstalk between lipid metabolism and immunity in dairy cows receiving essential fatty acids and conjugated linoleic acid.

Veshkini Arash A Hammon Harald M HM Vogel Laura L Viala Didier D Delosière Mylène M Tröscher Arnulf A Déjean Sébastien S Ceciliani Fabrizio F Sauerwein Helga H Bonnet Muriel M

Scientific reports 20220405 1

Essential fatty acids (EFA) and conjugated linoleic acids (CLA) are unsaturated fatty acids with immune-modulatory effects, yet their synergistic effect is poorly understood in dairy cows. This study aimed at identifying differentially abundant proteins (DAP) and their associated pathways in dairy cows supplied with a combination of EFA and CLA during the transition from antepartum (AP) to early postpartum (PP). Sixteen Holstein cows were abomasally infused with coconut oil as a control (CTRL) o ...[more]

PMID: 35383209

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Project description:The current study was designed to investigate the circulating miRNA expression signature of dairy cows during several key time points around calving, and under different fatty acids (FA) treatments to get insights into different aspects of metabolic adaptations and their interaction with administrated FA. In this trial, 32 transition Holstein dairy cows were randomly assigned to one of the following treatments: 1-CTRL (n = 8; coconut oil, Bio-Kokosöl #665, Kräuterhaus Sanct Bernhard, KG, Bad Ditzenbach, Germany; 38 and 76 g/d in antepartum (AP) and postpartum (PP)), 2- Essential fatty acids (EFA, n=8, a combination of linseed oil (DERBY® Leinöl #4026921003087, DERBY Spezialfutter GmbH, Münster, Germany; 39 and 78 g/d in AP and PP) and safflower oil (GEFRO Distelöl, GEFRO Reformversand Frommlet KG, Memmingen, Germany; 2 and 4 g/d in AP and PP), 3- conjugated linoleic acid (CLA, n = 8, Lutalin® ; cis-9, trans-11, 10 g/d trans- 10, cis-12 CLA, BASF SE, Lampertheim, Germany; 19 and 38 g/d in AP and PP), and 4- EFA+CLA, a combination of linseed oil, safflower oil and Lutalin® from day 63 AP to day 63 PP. From each cow, four blood samples were collected individually on days -21, +1, +28, and +63 relative to calving, and the plasma fraction was used for RNA extraction. Total RNA was extracted from 128 plasma samples using the QIAGEN miRNeasy serum/plasma kit (Qiagen GmbH, Hilden, Germany), and checked for yield and purity using the small RNA kit on a 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). We used the QIAseq miRNA library kit (QIAGEN, Hilden, Germany) for library preparation and Caliper (PerkinElmer, Waltham, MA) for quality check. NGS was performed on the libraries using the NovaSeq 6000 (Illumina, San Diego, CA) setting on single-end 100 bp mode on the respective platform of IGA Technology Services, Udine, Italy (https://igatechnology.com/). Raw-sequencing data was checked for quality (format conversion to FASTQ, demultiplexing, Adapter Trimming, and UMI Remova) using the Illumina Bcl2fastq2 Conversion Software (version 2.20). Detected sequences were analyzed with the miRDeep2 (miRDeep2.pl) software package (version 2.0.0.5) to detect known miRNA and predict putative novel miRNA (using human, ovine, and goat miRNA datasets). MiRDeep2 was fed with the Bos taurus miRNA collection available at miRBase (www.mirbase.org). Read counts for each known and novel miRNA were compiled using the HTSeq-count script. The quantified counts were normalized using the TMM method via the edgeR (version 3.12.0) package in R software (Version 4.0). Only miRNA with at least one count per million over at least 2 samples were taken into consideration for the analysis. Differentially expressed miRNA (DEM) during the time points were identified by performing generalized linear model (GLM) likelihood ratio tests (glmLRT) using the GLM approach in edgeR. The DEMs have been interpreted separately I-during different time points, and also II-between treatments, to gain insight into metabolic and immune adaptation, as well as the role of fatty acids at each time point. A time-affected DEM identified key signaling pathways that are involved in initiating and regulating metabolic shifts during the transition from gestation (non-lactation) to lactation. Although, fatty acid treatments had only minor effects on miRNA expression (insufficient DEM to perform pathway analysis).

Project description:The objective of this study was to determine whether adding phytoncide oil (PO) and soybean oil (SBO) to the dairy cow diet could increase milk conjugated linoleic acid (CLA) and depress methane (CH4) emissions in Holstein dairy cows. Rumen fermentation was conducted at four levels of SBO (0, 1, 2, and 4%, on DM basis) and two levels of PO (0 and 0.1%, on DM basis) with in vitro experiment. To evaluate blood parameters, fecal microbe population, milk yield and fatty acid compositions, and CH4 production, in vivo experiment was conducted using 38 Holstein dairy cows divided into two groups of control (fed TMR) and treatment (fed TMR with 0.1% PO and 2% SBO as DM basis). In the in vitro study (Experiment 1), PO or SBO did not affect rumen pH. However, SBO tended to decrease ruminal ammonia-N (p = 0.099). Additionally, PO or SBO significantly decreased total gas production (p = 0.041 and p = 0.034, respectively). Both PO and SBO significantly decreased CH4 production (p < 0.05). In addition, PO significantly increased both CLA isomers (c9, t11 and t10, c12 CLA) (p < 0.001). Collectively, 0.1% PO and 2% SBO were selected resulting in most effectively improved CLA and decreased CH4 production. In the in vivo study (Experiment 2), 0.1% PO with 2% SBO (PSO) did not affect complete blood count. However, it decreased blood urea nitrogen and magnesium levels in blood (p = 0.021 and p = 0.01, respectively). PSO treatment decreased pathogenic microbes (p < 0.05). It increased milk yield (p = 0.017) but decreased percentage of milk fat (p = 0.013) and MUN level (p < 0.01). In addition, PSO treatment increased both the concentration of CLA and PUFA in milk fat (p < 0.01). Finally, it decreased CH4 emissions from dairy cows. These results provide compelling evidence that a diet supplemented with PSO can simultaneously increase CLA concentration and decrease CH4 production with no influence on the amount of milk fat (kg/day) in Holstein dairy cows.

Project description:Phosphoproteomics is a cutting-edge technique that can be utilized to explore adipose tissue (AT) metabolism by quantifying the repertoire of phospho-peptides (PP) in AT. Dairy cows were supplemented with conjugated linoleic acid (CLA, n = 5) or a control diet (CON, n = 5) from 63 d prepartum to 63 d postpartum; cows were slaughtered at 63 d postpartum and AT was collected. We performed a quantitative phosphoproteomics analysis of subcutaneous (SC) and omental (OM) AT using nanoUPLC-MS/MS and examined the effects of CLA supplementation on the change in the phosphoproteome. A total of 5919 PP were detected in AT, and the abundance of 854 (14.4%) were differential between CON and CLA AT (p ≤ 0.05 and fold change ± 1.5). The abundance of 470 PP (7.9%) differed between OM and SC AT, and the interaction treatment vs. AT depot was significant for 205 PP (3.5% of total PP). The integrated phosphoproteome demonstrated the up- and downregulation of PP from proteins related to lipolysis and lipogenesis, and phosphorylation events in multiple pathways, including the regulation of lipolysis in adipocytes, mTOR signaling, insulin signaling, AMPK signaling, and glycolysis. The differential regulation of phosphosite on a serine residue (S777) of fatty acid synthase (FASN) in AT of CLA-supplemented cows was related to lipogenesis and with more phosphorylation sites compared to acetyl-coenzyme A synthetase (ACSS2). Increased protein phosphorylation was seen in acetyl-CoA carboxylase 1 (ACACA;8 PP), FASN (9 PP), hormone sensitive lipase (LIPE;6 PP), perilipin (PLIN;3 PP), and diacylglycerol lipase alpha (DAGLA;1 PP) in CLA vs. CON AT. The relative gene expression in the SC and OM AT revealed an increase in LIPE and FASN in CLA compared to CON AT. In addition, the expression of DAGLA, which is a lipid metabolism enzyme related to the endocannabinoid system, was 1.6-fold higher in CLA vs. CON AT, and the expression of the cannabinoid receptor CNR1 was reduced in CLA vs. CON AT. Immunoblots of SC and OM AT showed an increased abundance of FASN and a lower abundance of CB1 in CLA vs. CON. This study presents a complete map of the SC and the OM AT phosphoproteome in dairy cows following CLA supplementation and discloses many unknown phosphorylation sites, suggestive of increased lipid turnover in AT, for further functional investigation.

Dataset Information

Plasma proteomics reveals crosstalk between lipid metabolism and immunity in dairy cows receiving essential fatty acids and conjugated linoleic acid.

Publications

Plasma proteomics reveals crosstalk between lipid metabolism and immunity in dairy cows receiving essential fatty acids and conjugated linoleic acid.

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