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Alanyl-glutamine but not glycyl-glutamine improved the proliferation of enterocytes as glutamine substitution in vitro

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Abstract

The synthetic dipeptides alanyl-glutamine (Ala-Gln) and glycyl-glutamine (Gly-Gln) are used as Gln substitution to provide energy source in the gastrointestinal tract due to their high solubility and stability. This study aimed to investigate the effects of Gln, Ala-Gln and Gly-Gln on mitochondrial respiration and protein turnover of enterocytes. Intestinal porcine epithelial cells (IPEC-J2) were cultured for 2 days in Dulbecco’s modified Eagle’s-F12 Ham medium (DMEM-F12) containing 2.5 mM Gln, Ala-Gln or Gly-Gln. Results from 5-ethynyl-2′-deoxyuridine incorporation and flow cytometry analysis indicated that there were no differences in proliferation between free Gln and Ala-Gln-treated cells, whereas Gly-Gln treatment inhibited the cell growth compared with Gln treatment. Significantly lower mRNA expressions of Sp1 and PepT1 were also observed in Gly-Gln-treated cells than that of Ala-Gln treatment. Ala-Gln treatment increased the basal respiration and ATP production, compared with free Gln and Gly-Gln treatments. There were no differences in protein turnover between free Gln and Ala-Gln-treated cells, but Gly-Gln treatment reduced protein synthesis and increased protein degradation. Ala-Gln treatment stimulated mTOR activation whereas Gly-Gln decreased mTOR phosphorylation and increased the UB protein expression compared with free Gln treatment. These results indicate that Ala-Gln has the very similar functional profile to free Gln in porcine enterocytes in vitro and can be substituted Gln as energy and protein sources in the gastrointestinal tract.

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Abbreviations

4EBP1:

4E binding protein-1

AA:

Amino acids

Ala-Gln:

Alanyl-glutamine

ATP:

Adenosine triphosphate

DMEM-F12:

Dulbecco’s modified Eagle’s-F12 Ham medium

ECAR:

Extracellular acidification rate

EdU:

5′-Ethynyl-2′-deoxyuridine

FCCP:

Carbonyl cyanide-p-trifluoromethoxyphenylhydrazone

Gln:

Glutamine

Gly-Gln:

Glycyl-glutamine

IPEC:

Intestinal porcine epithelial cell

mTOR:

Mammalian target of rapamycin

OCR:

Oxygen consumption rate

PepT1:

Peptide transporter 1

S6K1:

Ribosomal protein S6 kinase-1

Sp1:

Specificity protein 1

UB:

Ubiquitin

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Acknowledgements

This study was in part supported by the National Natural Science Foundation of China (Nos. 31330075, 31372326, 31672433, 31301989 and 31560640), Key Programs of frontier scientific research of the Chinese Academy of Sciences (QYZDY-SSW-SMC008) and National Basic Research Program of China (2013CB127302). We thank Changsha Lvye Biotechnology Limited Company Academician Expert Workstation, Guangdong Wangda Group Academician Workstation for Clean Feed Technology Research and Development in Swine, Guangdong Hinapharm Group Academician Workstation for Biological Feed and Feed Additives and Animal Intestinal Health for providing technical assistance.

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Authors and Affiliations

  1. National Engineering Laboratory for Pollution Control and Waste Utilization in Livestock and Poultry Production, Key Laboratory of Agro-ecological Processes in Subtropical Region, Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha, 410125, Hunan, China

    Bie Tan, Honglan Liu, Guangzu He, Hao Xiao & Yulong Yin

  2. College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha, 410128, Hunan, China

    Bie Tan, Guangzu He, Jianping Wu & Jun Fang

  3. College of Animal Science and Technology, Hunan Agricultural University, Changsha, 410128, Hunan, China

    Bie Tan, Dingfu Xiao & Yulong Yin

  4. Department of Animal Science, University of California, Davis, CA, 95616, USA

    Bie Tan & Yanhong Liu

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  1. Bie Tan
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  2. Honglan Liu
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  3. Guangzu He
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  4. Hao Xiao
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  5. Dingfu Xiao
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  6. Yanhong Liu
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  7. Jianping Wu
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  8. Jun Fang
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  9. Yulong Yin
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Correspondence to Jun Fang or Yulong Yin.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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All authors listed have contributed to conception, design, gathering, analysis or interpretation of data and have contributed to the writing and intellectual content of the article. All authors gave informed consent to the submission of this manuscript.

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Handling Editors: C.-A.A. Hu, Y. Yin, Y. Hou, G. Wu, Y. Teng.

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Tan, B., Liu, H., He, G. et al. Alanyl-glutamine but not glycyl-glutamine improved the proliferation of enterocytes as glutamine substitution in vitro. Amino Acids 49, 2023–2031 (2017). https://doi.org/10.1007/s00726-017-2460-z

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  • DOI: https://doi.org/10.1007/s00726-017-2460-z

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