title: Selectin inhibition modulates NF-kappa B and AP-1 signaling after liver ischemia/reperfusion.
abstract: The infiltration of neutrophils after ischemia and reperfusion (I/R) is facilitated by the expression of adhesion molecules on the surface of both leukocytes and endothelial cells. Adhesion molecules of the selectin family are of particular importance at the onset of neutrophil mediated injury, as demonstrated by the occurrence of many cellular interactions with the final extravasation of inflammatory leukocytes at the site of I/R damage. Previous studies demonstrated a prevention of neutrophil extravasation and protection of ischemic damage when a small anti-selectin molecule was used. In this study, we tested a new small anti-selectin compound (OC-229) in a murine model of partial hepatic I/R. The aim of this study was to determine the effect of OC-229 on liver function and histology after I/R and to evaluate its role in the modulation of the inflammatory molecular signaling pathways of NF-kappa B and AP-1 under the same experimental condition. Mice subjected to 90 min of partial (70-80%) hepatic ischemia and 3 h of reperfusion were divided into three groups (n = 9/group): sham, ischemic control, and treated group, which received 25 mg/kg of the anti-selectin small molecule OC-229. These groups were studied when the treatment was given at the time of reperfusion (no pretreatment was given). The parameters measured at 3 h of reperfusion included liver function tests (ALT and AST), liver histology, and liver tissue electrophoretic mobility shift assay (EMSA) for NF-kappa B and AP-1. It was demonstrated that the multiselectin inhibitor OC-229 offered significant protection for the ischemic liver when given at 25 mg/kg at the time of reperfusion. ALT and AST serum levels significantly decreased when the ischemic control and the group receiving OC-229 were compared (p = .01). Treated animals demonstrated better histological findings as well. The EMSA showed dissociation of NF-kappa B and AP-1 activity in the liver nuclear extracts after selectin inhibition treatment. A reduction in the activity of AP-1 and an increment in NF-kappa B activation was seen. In this work, we obtained evidence that the small-molecule selectin inhibitor OC-229 offered functional and histological protection of the ischemic liver when given at 25 mg/kg at the time for reperfusion. There was dissociation in the activation signals of NF-kappa B and AP-1. Increase in NF-kappa B and reduction of the activation of AP-1 were noted at 3 h of reperfusion.
authors.affiliation: Borgess Research Institute, Trauma, Surgery Research Sciences, and Molecular Biology, Michigan State University, East Lansing, Michigan, USA.
authors.forename: Luis H
authors.initials: LH
authors.lastname: Toledo-Pereyra
authors.forename: Fernando
authors.initials: F
authors.lastname: Lopez-Neblina
authors.forename: Alex B
authors.initials: AB
authors.lastname: Lentsch
authors.forename: Roberto
authors.initials: R
authors.lastname: Anaya-Prado
authors.forename: Suzanne J
authors.initials: SJ
authors.lastname: Romano
authors.forename: Peter A
authors.initials: PA
authors.lastname: Ward
mesh_terms: D000410:Alanine Transaminase / Q000097:blood; D000818:Animals; D001219:Aspartate Aminotransferases / Q000097:blood; D002934:Cinnamates / Q000494:pharmacology; D024202:Electrophoretic Mobility Shift Assay; D007093:Imidazoles / Q000494:pharmacology; D008099:Liver / Q000098:blood supply* / Q000187:drug effects / Q000293:injuries / Q000473:pathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D016328:NF-kappa B / Q000502:physiology*; D009504:Neutrophils / Q000502:physiology; D015427:Reperfusion Injury / Q000473:pathology / Q000503:physiopathology* / Q000517:prevention & control; D019087:Selectins / Q000187:drug effects* / Q000502:physiology; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D018808:Transcription Factor AP-1 / Q000502:physiology*
chemical_list: C514929:3-(4-(5-(4-(2-dihexylcarbamoylvinyl)phenyl)-1H-imidazol-4-yl)phenyl)acrylic acid; D002934:Cinnamates; D007093:Imidazoles; D016328:NF-kappa B; D019087:Selectins; D018808:Transcription Factor AP-1; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase