title: Biomarkers of oxidative damage to predict ischaemia-reperfusion injury in an isolated organ perfusion model of the transplanted kidney.
abstract: Ischaemia-reperfusion (IR) injury is known to be a risk factor influencing both short and long-term graft function following transplantation. The pathophysiology of IR injury is suggested to involve elevated reactive oxygen species production resulting in oxidative damaged cellular macromolecules. The objective of this study was to evaluate oxidative damage following IR using an isolated organ perfusion model of the transplanted kidney, in order to determine a simple, preferably non-invasive biomarker for IR injury. Porcine kidneys were retrieved with 10 or 40 min warm ischaemic (WI) time and haemoperfused for 6 h on an isolated organ perfusion machine. ELISA was used to detect carbonyls, 8-isporostane and 8-hydroxy-2'-deoxyguanosine, representing protein, lipid and DNA damage respectively in pre and post reperfusion samples of plasma, urine and biopsy material. Plasma carbonyl and 8-isporostane and were significantly increased in the 40 min group compared to pre-perfusion (0.96 +/- 0.10 vs. 0.62 +/- 0.06, P < 0.001 and 1.57(1.28-4.9) vs. 0.36(0.09-0.59), P < 0.05). The levels also correlated with creatinine clearance used to determine renal function (r = - 0.6150, P < 0.01 and r = - 0.7727, P < 0.01). The results of this study suggest both plasma carbonyl and 8-isporostane to be reliable biomarkers to predict the level IR injury.
authors.affiliation: Transplant Surgery Group, Department of Cardiovascular Sciences, University of Leicester, Leicester, UK. hlw8@le.ac.uk
authors.forename: Helen L
authors.initials: HL
authors.lastname: Waller
authors.forename: Simon J F
authors.initials: SJ
authors.lastname: Harper
authors.forename: Sarah A
authors.initials: SA
authors.lastname: Hosgood
authors.forename: Atul
authors.initials: A
authors.lastname: Bagul
authors.forename: Bin
authors.initials: B
authors.lastname: Yang
authors.forename: Mark D
authors.initials: MD
authors.lastname: Kay
authors.forename: Monika
authors.initials: M
authors.lastname: Kaushik
authors.forename: Michael L
authors.initials: ML
authors.lastname: Nicholson
mesh_terms: D000818:Animals; D019540:Area Under Curve; D015415:Biomarkers / Q000737:chemistry; D002244:Carbon / Q000378:metabolism; D004249:DNA Damage; D015237:Dinoprost / Q000031:analogs & derivatives / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D007668:Kidney / Q000473:pathology; D016030:Kidney Transplantation / Q000379:methods*; D015227:Lipid Peroxidation; D018384:Oxidative Stress; D010477:Perfusion; D017382:Reactive Oxygen Species; D015427:Reperfusion Injury / Q000175:diagnosis* / Q000473:pathology; D013552:Swine
chemical_list: D015415:Biomarkers; D017382:Reactive Oxygen Species; C075750:8-epi-prostaglandin F2alpha; D002244:Carbon; D015237:Dinoprost