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Alpha-ketoglutarate inhibits glutamine degradation and enhances protein synthesis in intestinal porcine epithelial cells

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Abstract

α-Ketoglutarate (AKG) is a key intermediate in glutamine metabolism. Emerging evidence shows beneficial effects of AKG on clinical and experimental nutrition, particularly with respect to intestinal growth and integrity. However, the underlying mechanisms are unknown. Intestinal porcine epithelial cells (IPEC-1) were used to test the hypothesis that AKG inhibits glutamine degradation and enhances protein synthesis. IPEC-1 cells were cultured for 3 days in Dulbecco’s modified Eagle’s-F12 Ham medium (DMEM-F12) containing 0, 0.2, 0.5 or 2 mM of AKG. At the end of the 3-day culture, cells were used to determine l-[U-14C]glutamine utilization, protein concentration, protein synthesis, and the total and phosphorylated levels of the mammalian target of the rapamycin (mTOR), ribosomal protein S6 kinase-1 (S6K1) and eukaryotic initiation factor (eIF) 4E-binding protein-1 (4E-BP1). Compared with 0 mM of AKG (control), 0.2 and 0.5 mM of AKG dose-dependently reduced (P < 0.05) glutamine degradation and the production of glutamate, alanine and aspartate in IPEC-1 cells. Addition of 0.5 and 2 mM of AKG to culture medium enhanced protein synthesis (P < 0.05) by 78 and 101% without affecting protein degradation, compared to the control group. Rapamycin (50 nM; a potent inhibitor of mTOR) attenuated the stimulatory effect of AKG on protein synthesis. Consistent with these metabolic data, the addition of 0.5 or 2 mM of AKG to culture medium increased (P < 0.05) the phosphorylated levels of mTOR, S6k1 and 4E-BP1 proteins. Collectively, these results indicate that AKG can spare glutamine and activate the mTOR signaling pathway to stimulate protein synthesis in intestinal epithelial cells.

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Abbreviations

AKG:

α-Ketoglutarate

BSA:

Bovine serum albumin

DMEM-F12:

Dulbecco’s modified Eagle’s F12 Ham medium

FBS:

Fetal bovine serum

IPEC-1:

Intestinal porcine epithelial cells-1

mTOR:

Mammalian target of rapamycin

4E-BP1:

4E-Binding protein-1

S6K1:

70-kDa Ribosomal protein S6 kinase-1

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Acknowledgments

This research was jointly supported by the National Natural Science Foundation of China (30901041, 30901040, 30928018, 30972156, 30871801, 30828024, 30828025, 30771558 and 30700581), National 863 Project (2008AA10Z316), National Basic Research Project (2009CB118800), the CAS/SAFEA International Partnership Program for Creative Research Teams, the Program for Innovative Research Groups of Hubei Provincial Natural Science Foundation (Grant No. 2007ABC009), Texas AgriLife Research, American Heart Association (10GRNT4480020) and the Thousand-People-Talent program at the China Agricultural University.

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Authors and Affiliations

  1. Research Center for Healthy Breeding of Livestock and Poultry and Key Laboratory for Agro-ecological Processes in Subtropical Region, Institute of Subtropical Agriculture, The Chinese Academy of Sciences, Hunan, 410125, China

    Kang Yao, Yulong Yin & Guoyao Wu

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

    Kang Yao, Xilong Li, Pengbin Xi, Junjun Wang, Jian Lei & Guoyao Wu

  3. Ministry of Agriculture Key Laboratory of Animal Nutrition and Feed Science (South China), Institute of Animal Science, Guangdong Academy of Agricultural Sciences, Wushan Road, Tianhe District, Guangzhou, 510640, Guangdong, People’s Republic of China

    Pengbin Xi

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

    Junjun Wang & Guoyao Wu

  5. Hubei Key Laboratory of Animal Nutrition and Feed Science, Wuhan Polytechnic University, Wuhan, 430023, China

    Yongqing Hou

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  1. Kang Yao
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  2. Yulong Yin
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Correspondence to Kang Yao, Yulong Yin or Guoyao Wu.

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Yao, K., Yin, Y., Li, X. et al. Alpha-ketoglutarate inhibits glutamine degradation and enhances protein synthesis in intestinal porcine epithelial cells. Amino Acids 42, 2491–2500 (2012). https://doi.org/10.1007/s00726-011-1060-6

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  • DOI: https://doi.org/10.1007/s00726-011-1060-6

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