Contractile Responsiveness of Ventricular Myocytes to Isoproterenol Is Regulated by Induction of Nitric Oxide Synthase Activity in Cardiac Microvascular Endothelial Cells in Heterotypic Primary Culture

• Published in: Circulation research 1995-09, Vol.77 (3), p.486-493
• Main Authors: Ungureanu-Longrois, Dan, Balligand, Jean-Luc, Okada, Ikutaro, Simmons, William W, Kobzik, Lester, Lowenstein, Charles J, Kunkel, Steven L, Michel, Thomas, Kelly, Ralph A, Smith, Thomas W
• Format: Article
• Language: English
• Subjects: Amino Acid Oxidoreductases - biosynthesis; Animals; Cells, Cultured; Endothelium, Vascular - enzymology; Heart Ventricles; Interleukin-1 - pharmacology; Isoproterenol - pharmacology; Lipopolysaccharides - pharmacology; Male; Myocardial Contraction - drug effects; Nitric Oxide - physiology; Nitric Oxide Synthase; Rats; Rats, Sprague-Dawley; Transforming Growth Factor beta - pharmacology
• ISSN: 0009-7330; 1524-4571
• DOI: 10.1161/01.RES.77.3.486

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-alpha (TNF-alpha) antiserum. Interferon-gamma (IFN-gamma), which by itself does not induce iNOS in CMEC, potentiates and accelerates iNOS induction by IL-1 beta. Transforming growth factor-beta (TGF-beta) decreases iNOS activity, protein content, and mRNA abundance in IL-1 beta- and IFN-gamma-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-1 beta, a cytokine that alone does not affect myocyte contractile function in vitro. Baseline contractile amplitude, at 2 Hz and 37 degrees C, of myocytes in heterotypic culture with IL-1 beta-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 N-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-beta release and activation could act to limit the extent of endothelial cell iNOS induction in vivo.(Circ Res. 1995;77:486-493.)