Substitution of Dietary Sulfur Amino Acids by dl-2-Hydroxy-4-Methylthiobutyric Acid Reduces Fractional Glutathione Synthesis in Weaned Piglets

J Nutr. 2020 Apr 1;150(4):722-729. doi: 10.1093/jn/nxz272.

Abstract

Background: Cys is limiting for reduced glutathione (GSH) synthesis and can be synthesized from Met. We hypothesized that the dietary Met hydroxyl analogue dl-2-hydroxy-4-methylthiobutyric acid (dl-HMTBA) affects Cys and GSH metabolism and oxidative stress defense differently than Met.

Objective: The objective was to elucidate whether dl-HMTBA supplementation of a Met-deficient diet affects Cys flux, GSH fractional synthetic rate (FSR), and the basal oxidative stress level relative to Met supplementation in pigs.

Methods: Twenty-nine male German Landrace piglets aged 28 d were allocated to 3 dietary groups: a basal diet limiting in Met (69% of Met plus Cys requirement) supplemented with either 0.15% l-Met (LMET; n = 9), 0.15% dl-Met (DLMET; n = 11), or 0.17% dl-HMTBA (DLHMTBA; n = 9) on an equimolar basis. At age 54 d the pigs received a continuous infusion of [1-13C]-Cys to calculate Cys flux and Cys oxidation. After 3 d, GSH FSR was determined by [2,2-2H2]-glycine infusion, and RBC GSH and oxidized GSH concentrations were measured. At age 62 d the animals were killed to determine hepatic mRNA abundances of enzymes involved in GSH metabolism, GSH concentrations, and plasma oxidative stress defense markers.

Results: The Cys oxidation was 21-39% and Cys flux 5-15% higher in the fed relative to the feed-deprived state (P < 0.001). On average, GSH FSR was 49% lower (P < 0.01), and RBC GSH and total GSH concentrations were 12% and 9% lower, respectively, in DLHMTBA and DLMET relative to LMET pigs (P < 0.05). In the feed-deprived state, Gly flux, the GSH:oxidized glutathione (GSSG) ratio, RBC GSSG concentrations, plasma oxidative stress markers, and the hepatic GSH content did not differ between groups.

Conclusions: Although GSH FSR was higher in LMET compared with DLMET or DLHMTBA feed-deprived pigs, these differences were not reflected by lower oxidative stress markers and antioxidant defense enzymes in LMET pigs.

Keywords: dl-2-hydroxy-4-methylthiobutyric acid; cysteine; fluxes; glutathione synthesis; glycine; pigs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / blood
  • Amino Acids, Sulfur / administration & dosage*
  • Animals
  • Antioxidants / analysis
  • Biomarkers / blood
  • Cysteine / blood
  • Diet / veterinary*
  • Dietary Supplements
  • Erythrocytes / chemistry
  • Glutathione / analysis
  • Glutathione / biosynthesis*
  • Glutathione / blood
  • Glycine / blood
  • Liver / chemistry
  • Male
  • Methionine / administration & dosage
  • Methionine / analogs & derivatives*
  • Oxidative Stress / physiology
  • Sus scrofa / metabolism*
  • Weaning

Substances

  • Amino Acids
  • Amino Acids, Sulfur
  • Antioxidants
  • Biomarkers
  • alpha-hydroxy-gamma-methylmercaptobutyric acid
  • Methionine
  • Glutathione
  • Cysteine
  • Glycine