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l-Glutamine or l-alanyl-l-glutamine prevents oxidant- or endotoxin-induced death of neonatal enterocytes

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Abstract

This study tested the hypothesis that L-glutamine (Gln) or L-alanyl-L-glutamine (Ala-Gln) prevents oxidant- or endotoxin-induced death of neonatal enterocytes. Enterocytes of neonatal pigs rapidly hydrolyzed Ala-Gln and utilized Gln. To determine whether Gln or Ala-Gln has a cytoprotective effect, IPEC-1 cells were cultured for 24 h in Gln-free Dulbecco’s modified Eagle’s-F12 Ham medium containing 0, 0.5, 2.0 or 5.0 mM Gln or Ala-Gln, and 0, 0.5 mM H2O2 or 30 ng/ml lipopolysaccharide (LPS). Without Gln or Ala-Gln, H2O2- or LPS-treated cells exhibited almost complete death. Gln or Ala-Gln at 0.5, 2 and 5 mM dose-dependently reduced H2O2- or LPS-induced cell death by 14, 54 and 95%, respectively, whereas d-glutamine, alanine, glutamate, ornithine, proline, glucosamine or nucleosides had no effect. To evaluate the effectiveness of Gln or Ala-Gln in vivo, 7-day-old piglets received one-week oral administration of Gln or Ala-Gln (3.42 mmol/kg body weight) twice daily and then a single intraperitoneal injection of LPS (0.1 mg/kg body weight); piglets were euthanized in 24 and 48 h to analyze intestinal apoptotic proteins and morphology. Administration of Gln or Ala-Gln to LPS-challenged piglets increased Gln concentrations in small-intestinal lumen and plasma, reduced intestinal expression of Toll-like receptor-4, active caspase-3 and NFkB, ameliorated intestinal injury, decreased rectal temperature, and enhanced growth performance. These results demonstrate a protective effect of Gln or Ala-Gln against H2O2- or LPS-induced enterocyte death. The findings support addition of Gln or Ala-Gln to current Gln-free pediatric amino acid solutions to prevent intestinal oxidative injury and inflammatory disease in neonates.

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Abbreviations

AA:

Amino acid

Ala:

l-Alanine

Ala-Gln:

l-Alanyl-l-glutamine

DMEM-F12:

Dulbecco’s modified Eagle’s F12 Ham medium

FBS:

Fetal bovine serum

Gln:

l-Glutamine

IPEC-1:

Intestinal porcine epithelial cell line-1

LPS:

Lipopolysaccharide

MyD88:

Myeloid differentiation factor 88

NFkB:

Nuclear factor kappa-light-chain enhancer of activator B cells

TLR4:

Toll-like receptor 4

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Acknowledgments

We thank Scott Jobgen and Elaine Jobgen for technical support, as well as Frances Mutscher for assistance in manuscript preparation. This work was supported by the National Research Initiative Competitive Grant from the USDA Cooperative State Research, Education, and Extension Service (no. 2008-35206-18764), Texas AgriLife Research (H-8200), and National Natural Science Foundation of China (no. 30600434).

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Authors and Affiliations

  1. Department of Animal Science, Texas A&M University, College Station, TX, 77843, USA

    Tony E. Haynes, Peng Li, Xilong Li, Junjun Wang, Darrell A. Knabe & Guoyao Wu

  2. Ajinomoto Co., Inc., Tokyo, 104-8315, Japan

    Kazuhiro Shimotori & Hiroyuki Sato

  3. Department of Chemistry and Biochemistry, Angelo State University, San Angelo, TX, 76909, USA

    Nick E. Flynn

  4. State Key Laboratory of Animal Nutrition, China Agricultural University, 100193, Beijing, China

    Junjun Wang & Guoyao Wu

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  1. Tony E. Haynes
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  2. Peng Li
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  3. Xilong Li
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  9. Guoyao Wu
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Correspondence to Guoyao Wu.

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Unless indicated, amino acids used and analyzed in this study are l-isomers.

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Haynes, T.E., Li, P., Li, X. et al. l-Glutamine or l-alanyl-l-glutamine prevents oxidant- or endotoxin-induced death of neonatal enterocytes. Amino Acids 37, 131–142 (2009). https://doi.org/10.1007/s00726-009-0243-x

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