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Proteomic analysis of plasma after branched chain enriched mixture supplementation in mice.

ABSTRACT: BACKGROUND: Branched chain amino acid (BCAA) supplementation is a recently identified strategy to promote longevity in mice. A proteomic approach was used to identify proteins which are differentially expressed in the sera of mice following supplementation with selected branched chain amino acid enriched mixture (BCAAem). FINDINGS: 12 male mice (C57Bl6, 9 months-old) were randomly assigned to unsupplemented (Control, n?=?6) and supplemented (BCAA, n?=?6, 0.1 mg/gr/day in drink water for 4 weeks). At the end of treatment total plasma samples from Control and BCAAem mice were separated by two-dimensional gel electrophoresis (2-DE). After staining, the gels were imaged and differential protein expression patterns were interrogated using image analysis software. Spots showing a different expression level were identified through a comparison with 2D maps found in databases officially recognized (ExPASy).Master gels of Control and BCAA mice exhibited slightly different 2-DE patterns as only 10 spots out of 500 appeared differentially expressed: 8 were upregulated (corresponding to Apolipoprotein A-I (APOA1), Complement factor B, Complement C3, Immunoglobulin light chain) and 2 appeared downregulated (Alpha-1-antitrypsin and unknown). CONCLUSIONS: Supplementation with BCAAem in mice results in a slight perturbation of the host serum proteome. Of particular interest is the increased Apolipoprotein A-I (APOAI) following treatment.

SUBMITTER: Brocca L

PROVIDER: S-EPMC3618008 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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Proteomic analysis of plasma after branched chain enriched mixture supplementation in mice.

Brocca Lorenza L Mascaro Anna A D'Antona Giuseppe G

Journal of the International Society of Sports Nutrition 20130403 1

<h4>Background</h4>Branched chain amino acid (BCAA) supplementation is a recently identified strategy to promote longevity in mice. A proteomic approach was used to identify proteins which are differentially expressed in the sera of mice following supplementation with selected branched chain amino acid enriched mixture (BCAAem).<h4>Findings</h4>12 male mice (C57Bl6, 9 months-old) were randomly assigned to unsupplemented (Control, n = 6) and supplemented (BCAA, n = 6, 0.1 mg/gr/day in drink water ...[more]

PMID: 23552612

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Project description:Skeletal muscle loss (sarcopenia) is a major clinical complication in alcoholic cirrhosis with no effective therapy. Skeletal muscle autophagic proteolysis and myostatin expression (inhibitor of protein synthesis) are increased in cirrhosis and believed to contribute to anabolic resistance. A prospective study was performed to determine the mechanisms of sarcopenia in alcoholic cirrhosis and potential reversal by leucine. In six well-compensated, stable, alcoholic patients with cirrhosis and eight controls, serial vastus lateralis muscle biopsies were obtained before and 7 hours after a single oral branched chain amino acid mixture enriched with leucine (BCAA/LEU). Primed-constant infusion of l-[ring-(2) H5 ]-phenylalanine was used to quantify whole-body protein breakdown and muscle protein fractional synthesis rate using liquid chromatography/mass spectrometry. Muscle expression of myostatin, mammalian target of rapamycin (mTOR) targets, autophagy markers, protein ubiquitination, and the intracellular amino acid deficiency sensor general control of nutrition 2 were quantified by immunoblots and the leucine exchanger (SLC7A5) and glutamine transporter (SLC38A2), by real-time polymerase chain reaction. Following oral administration, plasma BCAA concentrations showed a similar increase in patients with cirrhosis and controls. Skeletal muscle fractional synthesis rate was 9.63?±?0.36%/hour in controls and 9.05?±?0.68%/hour in patients with cirrhosis (P?=?0.54). Elevated whole-body protein breakdown in patients with cirrhosis was reduced with BCAA/LEU (P?=?0.01). Fasting skeletal muscle molecular markers showed increased myostatin expression, impaired mTOR signaling, and increased autophagy in patients with cirrhosis compared to controls (P?<?0.01). The BCAA/LEU supplement did not alter myostatin expression, but mTOR signaling, autophagy measures, and general control of nutrition 2 activation were consistently reversed in cirrhotic muscle (P?<?0.01). Expression of SLC7A5 was higher in the basal state in patients with cirrhosis than controls (P?<?0.05) but increased with BCAA/LEU only in controls (P?<?0.001).Impaired mTOR1 signaling and increased autophagy in skeletal muscle of patients with alcoholic cirrhosis is acutely reversed by BCAA/LEU.

Project description:AIMS/HYPOTHESIS:Branched-chain amino acids (BCAAs) and aromatic amino acids (AAAs) are associated with type 2 diabetes. However, repeated measurements of BCAA/AAA and their interactions with dietary interventions have not been evaluated. We investigated the associations between baseline and changes at 1 year in BCAA/AAA with type 2 diabetes in the context of a Mediterranean diet (MedDiet) trial. METHODS:We included 251 participants with incident type 2 diabetes and a random sample of 694 participants (641 participants without type 2 diabetes and 53 overlapping cases) in a case-cohort study nested within the PREvención con DIeta MEDiterránea (PREDIMED) trial. Participants were randomised to a MedDiet+extra-virgin olive oil (n = 273), a MedDiet+nuts (n = 324) or a control diet (n = 295). We used LC-MS/MS to measure plasma levels of amino acids. Type 2 diabetes was a pre-specified secondary outcome of the PREDIMED trial. RESULTS:Elevated plasma levels of individual BCAAs/AAAs were associated with higher type 2 diabetes risk after a median follow-up of 3.8 years: multivariable HR for the highest vs lowest quartile ranged from 1.32 for phenylalanine ([95% CI 0.90, 1.92], p for trend = 0.015) to 3.29 for leucine ([95% CI 2.03, 5.34], p for trend<0.001). Increases in BCAA score at 1 year were associated with higher type 2 diabetes risk in the control group with HR per SD = 1.61 (95% CI 1.02, 2.54), but not in the MedDiet groups (p for interaction <0.001). The MedDiet+extra-virgin olive oil significantly reduced BCAA levels after 1 year of intervention (p = 0.005 vs the control group). CONCLUSIONS/INTERPRETATION:Our results support that higher baseline BCAAs and their increases at 1 year were associated with higher type 2 diabetes risk. A Mediterranean diet rich in extra-virgin olive oil significantly reduced the levels of BCAA and attenuated the positive association between plasma BCAA levels and type 2 diabetes incidence. Clinical trial number: SRCTN35739639 ( www.controlled-trials.com ).

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Proteomic analysis of plasma after branched chain enriched mixture supplementation in mice.

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Proteomic analysis of plasma after branched chain enriched mixture supplementation in mice.

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