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Leucine stimulates ASCT2 amino acid transporter expression in porcine jejunal epithelial cell line (IPEC-J2) through PI3K/Akt/mTOR and ERK signaling pathways

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Abstract

Leucine has been shown to influence intestinal protein metabolism, cell proliferation and migration. Furthermore, our previous study demonstrated that branched-chain amino acids could modulate the intestinal amino acid and peptide transporters in vivo. As the possible mechanisms are still largely unknown, in the present work, we studied the transcriptional and translational regulation of leucine on amino acid transporter production in IPEC-J2 cells and the signaling pathways involved. Treatment of IPEC-J2 cells with 7.5 mM leucine enhanced the mRNA expression of the Na+-neutral AA exchanger 2 (ASCT2) and 4F2 heavy chain (4F2hc) and caused an increase in ASCT2 protein expression. Leucine also activated phosphorylation of 4E-BP1 and eIF4E through the phosphorylation of mTOR, Akt and ERK signaling pathways in IPEC-J2 cells. Pre-treatment of IPEC-J2 cells with inhibitors of mTOR and Akt (rapamycin and wortmannin) or an inhibitor of ERK (PD098059) for 30 min before leucine treatment attenuated the positive effect of leucine in enhancing the protein abundance of ASCT2. These results demonstrate that leucine could up-regulate the expression of the amino acid transporters (ASCT2) through transcriptional and translational regulation by ERK and PI3K/Akt/mTOR activation.

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Abbreviations

ASCT2:

Na+-neutral AA exchanger 2

BCAA:

Branched-chain amino acids

4F2hc:

4F2 heavy chain

LP:

Low protein

mTOR:

Mammalian target of rapamycin

RT-PCR:

Real-time polymerase chain reaction

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Acknowledgments

This research was supported by National Key Basic Research Program (2012CB124704 and 2013CB127305).

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All the authors declare that they have no competing interests in the present work.

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Authors and Affiliations

  1. State Key Laboratory of Animal Nutrition, Ministry of Agriculture Feed Industry Centre, College of Animal Science and Technology, China Agricultural University, Beijing, 100193, China

    Shihai Zhang, Man Ren, Xiangfang Zeng, Pingli He, Xi Ma & Shiyan Qiao

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  1. Shihai Zhang
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  2. Man Ren
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  3. Xiangfang Zeng
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Correspondence to Xiangfang Zeng.

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S. Zhang and M. Ren contributed equally to this work.

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Zhang, S., Ren, M., Zeng, X. et al. Leucine stimulates ASCT2 amino acid transporter expression in porcine jejunal epithelial cell line (IPEC-J2) through PI3K/Akt/mTOR and ERK signaling pathways. Amino Acids 46, 2633–2642 (2014). https://doi.org/10.1007/s00726-014-1809-9

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