© 1995 American Heart Association, Inc.
Articles |
Contractile Responsiveness of Ventricular Myocytes to Isoproterenol Is Regulated by Induction of Nitric Oxide Synthase Activityin Cardiac Microvascular EndothelialCells in Heterotypic Primary Culture
From the Cardiovascular Division, Department of Medicine, and Department of Pathology, Brigham and Women's Hospital and Harvard Medical School, and the Respiratory Physiology Program, Harvard School of Public Health (L.K.), Boston, Mass; the Department of Medicine, Johns Hopkins University, Baltimore, Md (C.J.L.); and the Department of Pathology, University of Michigan Medical School, Ann Arbor (S.L.K.).
Correspondence to Ralph A. Kelly, MD, Cardiovascular Division, Brigham and Women's Hospital, 75 Francis St, Boston, MA 02115.
Abstract Unlike large-vessel endothelial
cells in cell culture, cardiac microvascular
endothelial cells (CMEC) isolated from adult rat
ventricular muscle exhibit little detectable constitutive
nitric oxide (NO) synthase activity after isolation in vitro but
respond to specific combinations of inflammatory mediators with an
increase in inducible NO synthase (iNOS; type 2 NO synthase) activity.
CMEC iNOS is induced by soluble inflammatory mediators in
lipopolysaccharide-activated rat alveolar
macrophage–conditioned medium at 24 hours, and this induction
can be partially prevented by either interleukin-1 (IL-1) receptor
antagonist or a polyclonal anti–rat tumor necrosis
factor-
(TNF-) antiserum. Interferon-
(IFN-), which by
itself does not induce iNOS in CMEC, potentiates and accelerates iNOS
induction by IL-1ß. Transforming growth factor-ß (TGF-ß)
decreases iNOS activity, protein content, and mRNA abundance in
IL-1ß– and IFN-–pretreated CMEC. To determine whether NO
released by CMEC would affect myocyte contractile function in vitro,
freshly isolated ARVM were allowed to settle onto confluent,
serum-starved CMEC that had been pretreated for 24 hours with IL-1ß,
a cytokine that alone does not affect myocyte contractile
function in vitro. Baseline contractile amplitude, at 2 Hz and 37°C,
of myocytes in heterotypic culture with IL-1ß–pretreated CMEC was
not different from that of myocytes in control, homotypic myocyte
cultures. However, cocultured myocytes exhibited decreased contractile
responsiveness to 2 nmol/L isoproterenol compared with control cells,
and this could be reversed by the addition of 1 mmol/L
NG-monomethyl-L-arginine,
an inhibitor of NOS. Thus, activation of
endothelial cell iNOS by specific cytokines
affects the contractile responsiveness to isoproterenol of adjacent
cardiac myocytes in vitro. Reciprocal cell-cell signaling leading to
TGF-ß release and activation could act to limit the extent of
endothelial cell iNOS induction in vivo.
Key Words: interleukin-1 • interferon- • transforming growth factor-ß • interleukin-1 receptor antagonist
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