Proteomic analysis of rat livers in hepatic cold ischemia and reperfusion

CAMILA KNECHT; CECILIA, L. BALABAN; JOAQUIN V. RODRIGUEZ; EDGARDO E. GUIBERT; GERMÁN L. ROSANO

Rosario

Congreso; 8th Latin American Congress of Artificial Organs, Biomaterials and Tissue Engineering (VIII COLAOB); 2014

Sociedad Latinoamericana de Biomateriales, Órganos Artificiales e Ingeniería de Tejidos

Ischemia and reperfusion (IR) injury constitutes a pivotal mechanism of tissuedamage in pathological conditions such as stroke, myocardialinfarction, vascular surgery, and organ transplant.In particular hepatic IR is a major cause of liver damage during transplant, leading to itsfunctional deterioration and eventually to its rejection. Mantainingthe viability of the organ during its ischemic transfer from donor torecipient is mainly based on hypothermia, wich is applied to reducehepatic metabolic activity. Understanding how the rat liver adapts tocoldischemiamay aid in the developement of new clinical procedures aimed toimprove ischemic tissue protection.The present work deals with the setting of a rat model of liver donation to get samples for proteomic analysis. Livers were surgically removed and divided into four experimental groups (according to the stage of tissue sample collection): GI: Control - collection of sample immediately after surgical removal; GII: collection of sample after 90 min of ex vivo reperfusion using an isolated perfused rat liver model (IPRL); GIII:collection of sample after 24 hs of preservation in HTK (histidine-tryptophan-ketoglutaratebuffer) at 4 ºC and GIV: collection of sample after 24 hs of preservation in HTK at 4ºC plus 90 min of ex vivo reperfusion. These groups were chosen as representative steps in atypicall liver transplantation process..To identify differentially expressed proteins in hepatic IR samples versus control, we examined pooled liver samples from each experimental group and subjected them to two-dimensional gel electrophoresis. The first dimension was run in a Multiphor II apparatus (GE) under denaturing conditions. The second dimension was run vertically in a Protean II system (BioRad).Proteins were detected by Coomassie staining. The gels were scanned and analyzed with the ImageMaster software (GE). Selected spots weresubjected to MALDI-TOF for protein identification at the Analytical Biochemistry and Proteomics Unit (UByPA) of the Institute Pasteur in Montevideo.p { margin-bottom: 0.21cm; direction: ltr; color: #000000; orphans: 2; widows: 2; background: transparent }p.western { font-family: "Times New Roman", serif; font-size: 12pt; so-language: en-US }p.cjk { font-family: "Times New Roman", serif; font-size: 12pt; so-language: zh-CN }p.ctl { font-family: "Times New Roman", serif; font-size: 12pt; so-language: ar-SA }