Characterization of the cytoprotective action of peroxynitrite decomposition catalysts.
The formation of the powerful oxidant peroxynitrite (PN) from the reaction of superoxide anion with nitric oxide has been shown to be a kinetically favored reaction contributing to cellular injury and death at sites of tissue inflammation. The PN molecule is highly reactive causing lipid peroxidation as well as nitration of both free and protein-bound tyrosine. We present evidence for the pharmacological manipulation of PN with decomposition catalysts capable of converting it to nitrate. In target cells challenged with exogenously added synthetic PN, a series of metalloporphyrin catalysts (5,10,15,20-tetrakis(2,4,6-trimethyl-3, 3-disulfonatophenyl)porphyrinato iron (III) (FeTMPS); 5,10,15, 20-tetrakis(4-sulfonatophenyl)porphyrinato iron (III) (FeTPPS); 5,10, 15,20-tetrakis(N-methyl-4'-pyridyl)porphyrinato iron (III) (FeTMPyP)) provided protection against PN-mediated injury with EC50 values for each compound 30-50-fold below the final concentration of PN added. Cytoprotection was correlated with a reduction in the level of measurable nitrotyrosine. In addition, we found our catalysts to be cytoprotective against endogenously generated PN in endotoxin-stimulated RAW 264.7 cells as well as in dissociated cultures of hippocampal neurons and glia that had been exposed to cytokines. Our studies thus provide compelling evidence for the involvement of peroxynitrite in cytokine-mediated cellular injury and suggest the therapeutic potential of PN decomposition catalysts in reducing cellular damage at sites of inflammation.
Misko
T P
TP
Discovery Pharmacology, G. D. Searle, St. Louis, Missouri 63167, USA.
Highkin
M K
MK
Veenhuizen
A W
AW
Manning
P T
PT
Stern
M K
MK
Currie
M G
MG
Salvemini
D
D
D000818:Animals; D002384:Catalysis; D016923:Cell Death; D005290:Ferric Compounds / Q000378:metabolism*; D006801:Humans; D007249:Inflammation / Q000378:metabolism / Q000473:pathology; D008665:Metalloporphyrins / Q000378:metabolism*; D009566:Nitrates / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism; D011166:Porphyrins / Q000378:metabolism*; D051381:Rats; D014407:Tumor Cells, Cultured
C112917:5,10,15,20-tetrakis(2,4,6-trimethyl-3,3-disulfonatophenyl)porphyrinato iron(III); D005290:Ferric Compounds; D008665:Metalloporphyrins; D009566:Nitrates; D011166:Porphyrins; C092399:meso-tetrakis(1-methyl-4-pyridiniumyl)porphyrin; C039642:peroxynitric acid; D009569:Nitric Oxide; C016136:tetraphenylporphine sulfonate