Contribution of nitric oxide synthase to human neutrophil chemiluminescence

Luminescence. Nov-Dec 1999;14(6):335-9. doi: 10.1002/(SICI)1522-7243(199911/12)14:6<335::AID-BIO557>3.0.CO;2-#.

Abstract

A chemiluminescence (CL) assay has been used to measure the reactive oxygen species (ROS)-generating capacity of phagocytes. Primed neutrophils produce ROS and nitric oxide (NO) upon induction of nitric oxide synthase (NOS) activity. NO and superoxide (O(2)(-)) form peroxynitrite (ONOO(-)), and emit CL. We examined the involvement of NOS in the CL response of neutrophils using a method based on the modulation of enzyme activity of NOS by the substrate L-arginine and an inhibitor; L-NAME. We used lipopolysaccharide (LPS) as the neutrophil-priming agent. Addition of sodium azide (NaN(3)) with horseradish peroxidase (HRP) to luminol-dependent CL, gave a CL response that was significantly enhanced when 10 mmol/L L-arginine was present (p <0.05), suggesting that NOS activity contributed to the CL response of human neutrophils. LPS-primed luminol-dependent CL was significantly inhibited by L-NAME compared with D-NAME. The proportion of the difference between the two inhibitors in luminol-dependent CL was 12.3 +/- 15.0%. Therefore, approximately 12% of the LPS-primed luminol-dependent CL decrease induced by L-NAME indicated the contribution of NOS activity to the CL response.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Enzyme Inhibitors / pharmacology
  • Humans
  • Lipopolysaccharides / pharmacology
  • Luminescent Measurements
  • Luminol
  • NG-Nitroarginine Methyl Ester / pharmacology
  • Neutrophils / drug effects
  • Neutrophils / enzymology*
  • Nitric Oxide Synthase / antagonists & inhibitors
  • Nitric Oxide Synthase / chemistry
  • Nitric Oxide Synthase / metabolism*
  • Reactive Oxygen Species / metabolism

Substances

  • Enzyme Inhibitors
  • Lipopolysaccharides
  • Reactive Oxygen Species
  • Luminol
  • Nitric Oxide Synthase
  • NG-Nitroarginine Methyl Ester