Medical College of Wisconsin
CTSICores SearchResearch InformaticsREDCap

Treatment with {alpha}-phenyl-N-tert-butylnitrone, a free radical-trapping agent, abrogates inflammatory cytokine gene expression during alloimmune activation in rat cardiac allografts. J Pharmacol Exp Ther 2005 Feb;312(2):774-9

Date

09/07/2004

Pubmed ID

15347735

DOI

10.1124/jpet.104.076026

Scopus ID

2-s2.0-13244277595   5 Citations

Abstract

Spin-trapping nitrones such as alpha-phenyl-N-tert-butylnitrone (PBN) have traditionally been used to trap and stabilize free radicals for detection by electron paramagnetic resonance (EPR) spectroscopy. Unlike classical antioxidants, these agents have never been evaluated therapeutically in allograft transplantation. In the present study, we examined potential mechanisms of action of treatment with PBN in a rat model of acute cardiac allograft transplantation. Graft rejection was determined by histological examination and graft function determined by in situ sonomicrometry. DNA binding for nuclear factor (NF)-kappaB and activator protein (AP-1) were determined by gel shift assays. Western blot and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis was performed for inducible nitric-oxide synthase (iNOS) and inflammatory cytokines. Histological rejection scores were elevated in untreated allografts and decreased by treatment with PBN. In situ sonomicrometry revealed decreased heart rate and distended end diastolic and end systolic segment lengths with rejection. Although PBN did not alter heart rate, it did normalize the distention of both diastolic and systolic cardiac dimension. EPR spectroscopy revealed nitrosylation of myocardial heme protein in untreated allografts that was decreased by treatment with PBN. PBN also decreased iNOS protein and iNOS mRNA. RT-PCR analysis revealed enhanced cytokine gene expression for interferon-gamma, interleukin-6, and interleukin-10 in untreated allografts. Expression for these genes was potently inhibited or abolished in recipients treated with PBN. PBN treatment also decreased DNA binding of transcription factors, NF-kappaB and AP-1. Thus, PBN retains significant anti-inflammatory properties through its action to down-regulate cytokine gene expression that contribute to protection against acute alloimmune activation in cardiac allografts.

Author List

Pieper GM, Nilakantan V, Zhou X, Khanna AK, Johnson CP, Roza AM, Adams MB, Hilton G, Felix CC

Authors

Christopher P. Johnson MD Professor in the Surgery department at Medical College of Wisconsin
Allan M. Roza MD Professor in the Surgery department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Animals
Autoimmune Diseases
Blotting, Western
Cyclic N-Oxides
Cytokines
Down-Regulation
Electron Spin Resonance Spectroscopy
Electrophoretic Mobility Shift Assay
Free Radical Scavengers
Gene Expression
Graft Rejection
Heart Transplantation
NF-kappa B
Nitric Oxide Synthase
Nitric Oxide Synthase Type II
Nitrogen Oxides
Nuclear Proteins
Rats
Rats, Inbred Lew
Rats, Inbred WF
Transcription Factor AP-1
Transcription Factors
jenkins-FCD Prod-478 d1509cf07a111124a2d122fd3df854cc0b993c00