Contribution of nitric oxide synthase to human neutrophil chemiluminescence

✍ Scribed by Satoru Kudoh; Katsuhiko Suzuki; Mutsuo Yamada; Qiang Liu; Shigeyuki Nakaji; Kazuo Sugawara

• Publisher: John Wiley and Sons
• Year: 1999
• Tongue: English
• Weight: 71 KB
• Volume: 14
• Category: Article
• ISSN: 1522-7235
• DOI: 10.1002/(sici)1522-7243(199911/12)14:6<335::aid-bio557>3.0.co;2-#

✦ Synopsis

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 À ) form peroxynitrite (ONOO À ), 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 AE 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.