Novel synthesis of S-nitrosoglutathione and degradation by human neutrophils.
S-nitrosoglutathione (SNO-GSH), a stable derivative of nitric oxide, is an endothelium-derived relaxation factor, which provokes vasodilation, inhibits platelet aggregation, and inhibits neutrophil (PMN) superoxide anion (O2+) generation. We have established a novel method for synthesis of S-nitrosoglutathione using a column containing S-nitrosothiol covalently attached to agarose. S-nitrosoglutathione was a product as assessed after separation using C-18 reverse-phase HPLC and absorption spectroscopy. We examined the stability of SNO-GSH in the presence or absence of PMN. The half-life (mercuric acid diazotization) of SNO-GSH in Hepes was greater than 60 min. The addition of resting PMN did not affect the T1/2 of SNO-GSH. PMN exposed to N-fMet-Leu-Phe (FMLP, 10(-7) M) reduced measurable SNO-GSH (15 microM) at 5 min (48 +/- 5.0% control, P less than 0.05). Incubation (5 min, 37 degrees C) of PMN with 10 microM tenidap (an anti-inflammatory drug which inhibits PMN activation) before addition of FMLP blocked the PMN-dependent degradation of SNO-GSH (42 +/- 3 vs 78 +/- 1.3% control, P = 0.01). We confirmed the recovery of SNO-GSH through measurements by bioassay (platelet aggregation) and HPLC analysis. The degradation of S-nitrosothiols by activated neutrophils may reverse the inhibitory effect of S-nitrosothiols on PMN functions and contribute to tissue injury at sites of inflammation.
Clancy
R M
RM
Department of Medicine, New York University Medical Center, New York 10016.
Abramson
S B
SB
D003545:Cysteine / Q000737:chemistry; D005978:Glutathione / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007211:Indoles / Q000494:pharmacology; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism; D009603:Nitroso Compounds / Q000138:chemical synthesis* / Q000378:metabolism; D000078183:Oxindoles; D010894:Piroxicam / Q000494:pharmacology; D026422:S-Nitrosoglutathione; D012459:Salicylates / Q000494:pharmacology; D020156:Salicylic Acid
D007211:Indoles; D009603:Nitroso Compounds; D000078183:Oxindoles; D012459:Salicylates; D010894:Piroxicam; D009569:Nitric Oxide; D026422:S-Nitrosoglutathione; D009240:N-Formylmethionine Leucyl-Phenylalanine; C062866:tenidap; D005978:Glutathione; D003545:Cysteine; D020156:Salicylic Acid