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Molecular Pharmacology

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Rapid CommunicationAccelerated Communication

Induction of l-Arginine Transport Is Inhibited by Atrial Natriuretic Peptide: A Peptide Hormone as a Novel Regulator of Inducible Nitric-Oxide Synthase Substrate Availability

Alexandra K. Kiemer and Angelika M. Vollmar
Molecular Pharmacology September 2001, 60 (3) 421-426;
Alexandra K. Kiemer
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Angelika M. Vollmar
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Abstract

Background: The inducible nitric-oxide synthase (iNOS) synthesizes NO from l-arginine. Availability ofl-arginine is maintained by a lipopolysaccharide (LPS)-induced induction of the CAT-2B amino acids transporter. Recently, we could show that the cardiovascular hormone atrial natriuretic peptide (ANP) inhibits the induction of iNOS in LPS-stimulated macrophages via its guanylate cyclase-coupled A-receptor. Purpose: To investigate whether ANP exerts an effect on LPS-induced l-arginine uptake.Methods: Murine bone marrow derived macrophages were activated with LPS (1 μg/ml, 20 h) in the presence or absence of ANP or C-type natriuretic peptide (CNP). l-Arginine transport was determined by measuring the uptake ofl-[3H]arginine.l-[3H]Arginine influx was also determined in LPS-activated cells in the presence ofNG-monomethyl-l-arginine (l-NMMA), competitor amino acids, or ANP. Nitrite accumulation was determined in supernatants of LPS-activated cells cultured in the presence or absence ofl-ornithine. Results: ANP dose dependently (10−8–10−6M) inhibited LPS-inducedl-[3H]arginine uptake when added simultaneously with LPS, whereas it showed no effect when added simultaneously withl-[3H]arginine. The effect was abrogated by the A-receptor antagonist HS-142–1 (10 μg/ml). CNP (10−6 M) did not influencel-arginine transport. Competitor amino acids (10−2 M) inhibitedl-[3H]arginine uptake. An excess of unlabeled l-arginine (10−2 M) as well as its analogl-NMMA (10−3 M) also reduced l-[3H]arginine influx. l-Arginine uptake was critical for production of NO because l-ornithine significantly decreased LPS-induced nitrite accumulation.Conclusion: This work demonstrates that ANP inhibits LPS-induced l-arginine uptake via its guanylate cyclase-coupled A-receptor. Besides its influence on the induction of iNOS, this effect may represent an important and unique mechanism by which ANP regulates NO production in macrophages.

Footnotes

  • This work was supported by the Deutsche Forschungsgemeinschaft Vo 376/8–2. A.K.K. is supported by the “Bayerischer Habilitationsförderpreis”.

  • Abbreviations:
    iNOS
    inducible nitric-oxide synthase
    NOS
    nitric-oxide synthase
    ANP
    atrial natriuretic peptide
    LPS
    lipopolysaccharide
    NF-κB
    nuclear factor-κB
    NP
    natriuretic peptide
    NPR
    natriuretic peptide receptor
    CNP
    C-type natriuretic peptide
    BMM
    bone marrow macrophages
    PBS
    phosphate-buffered saline
    NMMA
    NG-monomethyl-l-arginine
    • Received February 9, 2001.
    • Accepted June 7, 2001.
  • The American Society for Pharmacology and Experimental Therapeutics
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Molecular Pharmacology: 60 (3)
Molecular Pharmacology
Vol. 60, Issue 3
1 Sep 2001
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Rapid CommunicationAccelerated Communication

Induction of l-Arginine Transport Is Inhibited by Atrial Natriuretic Peptide: A Peptide Hormone as a Novel Regulator of Inducible Nitric-Oxide Synthase Substrate Availability

Alexandra K. Kiemer and Angelika M. Vollmar
Molecular Pharmacology September 1, 2001, 60 (3) 421-426;

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Induction of l-Arginine Transport Is Inhibited by Atrial Natriuretic Peptide: A Peptide Hormone as a Novel Regulator of Inducible Nitric-Oxide Synthase Substrate Availability

Alexandra K. Kiemer and Angelika M. Vollmar
Molecular Pharmacology September 1, 2001, 60 (3) 421-426;
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