Regulatory Mechanism of Mammalian Target of Rapamycin Signaling Pathway on Arginine Transport in Porcine Intestinal Epithelial Cells

XIAO Hao; ZHAO Yi; WANG Li; TAN Bi'e

doi:10.3969/j.issn.1006-267x.2021.05.047

2021 , Vol. 33 >Issue 5: 2896 - 2906

DOI: https://doi.org/10.3969/j.issn.1006-267x.2021.05.047

Regulatory Mechanism of Mammalian Target of Rapamycin Signaling Pathway on Arginine Transport in Porcine Intestinal Epithelial Cells

XIAO Hao ,
ZHAO Yi ,
WANG Li ,
TAN Bi'e

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1. State Key Laboratory of Livestock and Poultry Breeding, Key Laboratory of Animal Nutrition and Feed Science in South China Ministry of Agriculture, Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Guangdong Key Laboratory of Animal Breeding and Nutrition, Institute of Animal Science, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China;
2. Observation and Experiment Station of Animal Nutrition and Feed Science in South-Central China, Ministry of Agriculture, Hunan Provincial Engineering Research Center for Healthy Livestock and Poultry Production, Key Laboratory of Agro-Ecological Processes in Subtropical Region, Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China

Received date: 2020-10-14

Online published: 2021-05-14

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Abstract

The purpose of this study was to investigate the mechanism of mammalian target of rapamycin (mTOR) signaling pathway regulating the proliferation of porcine intestinal epithelial cells and arginine (Arg) transport. Porcine intestinal epithelial cells (IPEC-J2 cells) were cultured in Dulbecco’s modified Eagle’s-high glucose Ham (DMEM-H) medium containing 100 or 350 μmol/L Arg, 0 or 10 nmol/L rapamycin (Rap) for 3 days, and then collected for the determination of cell proliferation, cell cycle, and the expression of related genes and protein of Arg transport, proliferation and apoptosis pathways. The results showed as follows:1) inhibition of mTOR signaling pathway by adding Rap could significantly reduce the cell number of G2-phase and S-phase (P<0.01), while elevating the Arg concentration could alleviate the inhibitory effect of Rap on cell proliferation. Rap reduced the cell apoptosis by activating the phosphatidylinositol-3-hydroxykinase (PI3K)-protein kinase (Akt)-B-cell lymphoma/leukemia-2 (Bcl2) signaling pathway. 2) Inhibition of mTOR signaling pathway by adding Rap could significantly increase the Arg uptake rate (P<0.01), and the mRNA and protein expression levels of cationic amino acid transporter 2 (CAT2) were significantly promoted after added Rap into the medium which containing 100 μmol/L Arg (P<0.01). It is further demonstrated that Rap might promote CAT2 expression through the protein kinase Cα (PKCα)-extracellular signal-regulated kinase (Erk)/cFos-CAT2 signaling pathway, thereby promoting Arg uptake. It is concluded that inhibition of mTOR signaling pathway by Rap can inhibit the proliferation of porcine intestinal epithelial cells, while increasing Arg concentration can effectively alleviate the inhibitory effects of Rap; Rap promotes Arg uptake by regulating PKCα-Erk/cFos-CAT2 signaling pathway in porcine intestinal epithelial cells, and increasing Arg concentration can promote the Arg uptake of cells. The results suggest that mTOR signaling pathway plays an important role in the utilization of Arg in porcine intestinal epithelial cells.

Key words： mTOR signaling pathway; arginine uptake; porcine intestinal epithelial cells; PKCα-Erk/cFos-CAT2 signaling pathway

Cite this article

XIAO Hao , ZHAO Yi , WANG Li , TAN Bi'e . Regulatory Mechanism of Mammalian Target of Rapamycin Signaling Pathway on Arginine Transport in Porcine Intestinal Epithelial Cells[J]. Chinese Journal of Animal Nutrition, 2021 , 33(5) : 2896 -2906 . DOI: 10.3969/j.issn.1006-267x.2021.05.047

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