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l-arginine stimulates CAT-1-mediated arginine uptake and regulation of inducible nitric oxide synthase for the growth of chick intestinal epithelial cells

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Abstract

l-arginine (l-Arg) uptake is mediated by members of cationic amino acid transporter (CAT) family and may coincide with the induction of nitric oxide synthases (NOS). The present study was conducted to investigate the extracellular concentrations of l-Arg regulating the CAT-1, CAT-4 and inducible NOS (iNOS) in chick intestinal epithelial cells. The cells were cultured for 4 days in Arg-free Dulbecco’s modified Eagle’s medium containing 10, 100, 200, 400, or 600 μM l-Arg. Cell viability, nitric oxide (NO) concentrations, uptake and metabolism of l-[3H]-Arg as well as expression of CAT-1, CAT-4, and iNOS were determined. Our results showed that l-Arg enhances cell growth with a maximal response at 10–400 μM. Addition of 100, 200, or 400 μM l-Arg increased the l-[3H]-Arg uptake, which was associated with greater conversion of l-[3H]-citrulline and NO production in comparison with 10 μM l-Arg group. Increasing extracellular concentrations of l-Arg from 10 to 400 μM dose dependently increased the levels of CAT-1 mRNA and protein, while no effect on CAT-4 mRNA abundance was found. Furthermore, supplementation of 100, 200, or 400 μM l-Arg upregulated the expression of iNOS mRNA, and the relative protein levels for iNOS in 200 and 400 μM l-Arg groups were higher than those in 10 and 100 μM l-Arg groups. Collectively, we conclude that the CAT-1 isoform plays a role in l-Arg uptake, and l-Arg-mediated elevation of NO via iNOS promotes the growth of chick intestinal epithelial cells.

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Acknowledgments

This research was supported by the earmarked fund for Modern Argo-Industry Technology Research System of China (No. CARS-41-K17) and the Program for Zhejiang Leading Team of S&T Innovation (No. 2011R50025-04).

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We declare that we have no conflict of interest.

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Authors and Affiliations

  1. Key Laboratory for Molecular Animal Nutrition of Ministry of Education, Feed Science Institute, College of Animal Science, Zhejiang University (Zijingang Campus), Hangzhou, 310058, China

    Chao Yuan, Xiaoyun Zhang, Qiang He, Junming Li, Jianjun Lu & Xiaoting Zou

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  1. Chao Yuan
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  2. Xiaoyun Zhang
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  3. Qiang He
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Correspondence to Jianjun Lu or Xiaoting Zou.

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Yuan, C., Zhang, X., He, Q. et al. l-arginine stimulates CAT-1-mediated arginine uptake and regulation of inducible nitric oxide synthase for the growth of chick intestinal epithelial cells. Mol Cell Biochem 399, 229–236 (2015). https://doi.org/10.1007/s11010-014-2249-2

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