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Production by the Atrial Natriuretic Peptide in Murine Macrophages1
*
Institute of Pharmacy, Center of Drug Research, Pharmaceutical Biology, University of Munich, Munich, Germany; and
Biochemical Pharmacology, University of Konstanz, Konstanz, Germany
The atrial natriuretic peptide (ANP) is suggested to regulate
inflammatory response by alteration of macrophage functions. The aim of
this study was to investigate whether ANP influences production of
TNF-
. TNF- production in murine bone marrow-derived macrophages
was induced by LPS, and TNF- secretion (±ANP) was determined by
L929 bioassay. ANP dose dependently
(10-8–10-6 M)
inhibited TNF- release by up to 95%. The effect was mediated via
the guanylate cyclase-coupled A receptor, as was shown by employing
dibutyryl-cGMP, the cGMP-inhibitory compound Ly-83583, and the A
receptor antagonist HS-142-1. A specific ligand of the natriuretic
peptide "clearance" receptor inhibited TNF- production only at
10-7 and 10-8 M, but
not at 10-6 M. The B receptor ligand C-type
natriuretic peptide showed no TNF--inhibitory effect. To investigate
the underlying mechanism of ANP-mediated TNF- inhibition, Northern
blot was performed. ANP-treated macrophages displayed decreased
TNF--mRNA levels. Besides the known inhibition of NF-
B
activation, in this study we demonstrated that ANP also attenuates the
activation of the proinflammatory transcription factor AP-1 (gel shift
assay). ANP did not alter subunit composition of AP-1 complexes, as was
shown by supershift assays applying anti-c-jun
and anti-c-fos Abs. To get information on the
ANP effect for human inflammatory processes, we investigated cytokine
production in human LPS-activated blood. ANP significantly attenuated
production of TNF- and IL-1ß without affecting production of IL-10
and IL-1ra. In summary, ANP was shown to attenuate TNF- production
of LPS-activated macrophages via cGMP. The inhibition is suggested to
involve transcriptional processes that are the result of reduced
activation of responsible transcription factors.
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