title: Plasma 3-nitrotyrosine is a biomarker in animal models of arthritis: Pharmacological dissection of iNOS' role in disease.
abstract: The contribution of inducible nitric oxide synthase (iNOS) to oxidative/nitrative stress is well-documented in inflammation, but difficult to quantify. Using a novel, recently developed assay for 3-nitrotyrosine (3-NT), we characterized iNOS activity and its inhibition in preclinical models of inflammation. In particular, we utilized the 3-NT assay to assess the role of iNOS in the disease pathology as well as for proof of pharmacology of iNOS inhibitors in an acute endotoxin challenge model, in models of rheumatoid arthritis (RA) such as rat adjuvant- and collagen-induced arthritis (AIA and CIA) and a model of osteoarthritis (OA) such as rat sodium monoiodoacetate-induced arthritis (MIA). Quantification of nitrotyrosine was performed using immuno-affinity 2-D LC-MS/MS assay. This assay is a very specific and reproducible and is amenable to a number of biological fluids. Plasma levels of 3-NT were significantly elevated in an acute model of inflammation (rat LPS) and in models of rheumatoid arthritis (adjuvant- and collagen-induced arthritis), and osteoarthritis (monoiodoacetate-induced arthritis). Plasma 3-NT correlated with the severity of the inflammatory response; thus, a 20-fold increase was observed in the rat LPS model, a 10-fold increase in AIA, and only a 2.5-fold elevation in CIA. Pharmacological intervention with iNOS inhibitors decreased 3-NT levels and associated pathology. 3-NT determination allowed for better elucidation of the role of iNOS in RA and OA disease pathology and provided proof of pharmacology for NOS inhibitors in animal models of RA and OA.
authors.affiliation: Pfizer Global Research and Development, St. Louis Laboratories, 700 Chesterfield Parkway, St. Louis, MO 63017, USA. olga.v.nemirovskiy@pfizer.com
authors.forename: Olga V
authors.initials: OV
authors.lastname: Nemirovskiy
authors.forename: Melissa R
authors.initials: MR
authors.lastname: Radabaugh
authors.forename: Poonam
authors.initials: P
authors.lastname: Aggarwal
authors.forename: Christie L
authors.initials: CL
authors.lastname: Funckes-Shippy
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Mnich
authors.forename: Debra M
authors.initials: DM
authors.lastname: Meyer
authors.forename: Teresa
authors.initials: T
authors.lastname: Sunyer
authors.forename: W
authors.initials: W
authors.lastname: Rodney Mathews
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Misko
mesh_terms: D000818:Animals; D001169:Arthritis, Experimental; D001172:Arthritis, Rheumatoid; D015415:Biomarkers / Q000097:blood; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007249:Inflammation; D052247:Nitric Oxide Synthase Type II / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000502:physiology*; D010003:Osteoarthritis; D051381:Rats; D012720:Severity of Illness Index; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000097:blood
chemical_list: D015415:Biomarkers; D004791:Enzyme Inhibitors; C002744:3-nitrotyrosine; D014443:Tyrosine; D052247:Nitric Oxide Synthase Type II