Monocyte-induced downregulation of nitric oxide synthase in cultured aortic endothelial cells

Nándor Marczin, Alexander Antonov, Andreas Papapetropoulos, David H. Munn, Renu Virmani, Frank D. Kolodgie, Ross Gerrity, John D. Catravas

Research output: Contribution to journalArticle

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Abstract

Since endothelium-dependent vasodilation is altered in atherosclerosis and enhanced monocyte/endothelial interactions are implicated in early atherosclerosis, we evaluated the effects of monocytes on the endothelial nitric oxide (NO) pathway by estimating release of biologically active NO from cultured endothelial cells and levels of constitutive NO synthase (ecNOS). NO release was estimated in a short-term bioassay using endothelial cell-induced cGMP accumulation in vascular smooth muscle (SM) cells. Exposure of SM cells to porcine aortic endothelial cells (PAECs) and human aortic endothelial cells (HAECs) produced large increases in SM cGMP content; this increase was prevented by N(G)-nitro-L-arginine methyl ester, the inhibitor of endothelial NOS. Confluent monolayers of PAECs and HAECs cocultured with monocytes also stimulated SM cGMP formation: however, NO release from these cultures was attenuated in a coculture time (2 to 48 hours)- and monocyte concentration (20 to 200x 103 per well)-dependent manner. This effect of monocyte adhesion appeared to be selective for NO release since other biochemical pathways, such as atriopeptin and isoproterenol-induced cyclic nucleotide accumulation within the endothelial cells, were not altered by monocytes. The effects of adherent monocytes on NO release were mimicked by monocyte-derived cytokines tumor necrosis factor (TNF)-α and interleukin (IL)-1α. Furthermore, the conditioned medium of monocytes contained significant quantities of these cytokines. Conditioned medium, as well as monocytes physically separated from the endothelial cells, attenuated NO release, suggesting that soluble factors may mediate the effects of monocytes. An IL-1β neutralizing antibody fully prevented the NO dysfunction in response to directly adherent monocytes. Superoxide dismutase, catalase, 4,5-dihydroxy-1,3-benzene disulfonic acid (Tiron), and exogenous L-arginine failed to improve NO release, suggesting that oxidant stress-induced inactivation of NO or limited substrate availability were not primarily responsible for the inhibiting effects of monocytes. Western blot analysis revealed reduced quantities of ecNOS in monocyte/endothelium cocultures, as well as in HAECs treated with monocyte-conditioned medium or TNF-α. Thus, adhesion of monocytes to endothelial cells and monocyte-derived secretory products downregulate steady state levels of ecNOS, an event associated with attenuated release of biologically active NO. This mechanism may potentially contribute to diminished endothelium-dependent and NO-mediated vasodilation in early atherosclerosis.

Original languageEnglish (US)
Pages (from-to)1095-1103
Number of pages9
JournalArteriosclerosis, thrombosis, and vascular biology
Volume16
Issue number9
DOIs
StatePublished - Jan 1 1996
Externally publishedYes

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Nitric Oxide Synthase
Monocytes
Down-Regulation
Endothelial Cells
Nitric Oxide
Conditioned Culture Medium
Endothelium
Atherosclerosis
Coculture Techniques
Interleukin-1
Vasodilation
Smooth Muscle Myocytes
Smooth Muscle
Swine
Tumor Necrosis Factor-alpha
1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium Salt
Cytokines
Cyclic Nucleotides
NG-Nitroarginine Methyl Ester
Atrial Natriuretic Factor

Keywords

  • atherosclerosis
  • cGMP
  • endothelium-derived relaxing factor
  • interleukin- 1
  • nitric oxide synthase

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

Cite this

Monocyte-induced downregulation of nitric oxide synthase in cultured aortic endothelial cells. / Marczin, Nándor; Antonov, Alexander; Papapetropoulos, Andreas; Munn, David H.; Virmani, Renu; Kolodgie, Frank D.; Gerrity, Ross; Catravas, John D.

In: Arteriosclerosis, thrombosis, and vascular biology, Vol. 16, No. 9, 01.01.1996, p. 1095-1103.

Research output: Contribution to journalArticle

Marczin, Nándor ; Antonov, Alexander ; Papapetropoulos, Andreas ; Munn, David H. ; Virmani, Renu ; Kolodgie, Frank D. ; Gerrity, Ross ; Catravas, John D. / Monocyte-induced downregulation of nitric oxide synthase in cultured aortic endothelial cells. In: Arteriosclerosis, thrombosis, and vascular biology. 1996 ; Vol. 16, No. 9. pp. 1095-1103.
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