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Peroxynitrite formation from activated human leukocytes

, : Peroxynitrite formation from activated human leukocytes. Biochemical and Biophysical Research Communications 224(2): 414-419

We showed direct evidence of peroxynitrite formation from polymorphonuclear cells (PMN) with the nitration of 4-hydroxyphenylacetic acid (HPA) to 4-hydroxy-3-nitrophenylacetic acid (NO-2HPA). Human PMN from healthy volunteers was stimulated with phorbol-12-myristate-13-acetate (PMA, 10 ng/ml) at 37 degree C in 2-(4-(2-hydroxyethyl)-1-piperazinyl)ethanesulfonic acid-buffered Hank's balanced salt solution (pH 7.4) with HPA (1 mM). NO-2HPA was detected under PMA stimulation only in the presence of myeloperoxidase inhibitor. NO-2HPA was eliminated by N-monomethyl-L-arginine (100 mu-M). The inhibition of myeloperoxidase appears to be essential to demonstrate the production of NO-2HPA since myeloperoxidase itself or its product, hypochlorite, reacted with peroxynitrite and hampered the formation of NO-2HPA.


PMID: 8702403

DOI: 10.1006/bbrc.1996.1041

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