Long-term cold storage preservation does not affect fatty livers from rats fed with a methionine and choline deficient diet

Abstract

Background Waiting lists that continue to grow and the lack of organs available for transplantation necessitate the use of marginal livers, such as fatty livers. Since steatotic livers are more susceptible to damage from ischemia and reperfusion, it was investigated whether fatty livers with different lipidomic profiles show a different outcome when subjected to long-term cold storage preservation. Methods Eight-week-old male Wistar rats fed for 2 weeks by a methionine-choline-deficient (MCD) diet or control diet were employed in this study. Livers were preserved in a University of Wisconsin (UW) solution at 4 °C for 6, 12 or 24 h and, after washout, reperfused for 2 h with a Krebs-Henseleit buffer at 37 °C. Hepatic enzyme release, bile production, O2-uptake, and portal venous pressure (PVP) were evaluated. The liver fatty acid profile was evaluated by a gas chromatography-mass spectrometry (GC/MS). Results MCD rats showed higher LDH and AST levels with respect to the control group. When comparing MCD livers preserved for 6, 12 or 24 h, no differences in enzyme release were found during both the washout or the reperfusion period. The same trend occurred for O2-uptake, PVP, and bile flow. A general decrease in SFA and MUFA, except for oleic acid, and a decrease in PUFA, except for arachidonic, eicosadienoic, and docosahexanaeoic acids, were found in MCD rats when compared with control rats. Moreover, the ratio between SFA and the various types of unsaturated fatty acids (UFA) was significantly lower in MCD rats. Conclusions Although prolonged cold ischemia negatively affects the graft outcome, our data suggest that the quality of lipid constituents could influence liver injury during cold storage: the lack of an increased hepatic injury in MCD may be justified by low SFA, which likely reduces the deleterious tendency toward lipid crystallization occurring under cold ischemia.

MCD Diet Modulates HuR and Oxidative Stress-Related HuR Targets in Rats

“…Five-week-old male Wistar rats (Charles River Laboratories, Calco, Italy) were used in this study. After a 1-week quarantine period, animals were fed with the MCD diet (Laboratorio Dottori Piccioni, Gessate, Italy) or isocaloric control diet (CTRL) for 3 weeks (3w; n = 7 for each group) or 6 weeks (6w; n = 15 MCD group; n = 6 CTRL group), as previously described [ 61 ]. At the end of diet treatment, rats were anesthetized with sodium pentobarbital injected intraperitoneally (40 mg/kg), the abdomen was opened, and blood was collected from the inferior caval vein.…”

MCD Diet Rat Model Induces Alterations in Zinc and Iron during NAFLD Progression from Steatosis to Steatohepatitis

“…Rats were anesthetized (Pentobarbital i.p. injection, 40 mg/kg), the livers were exposed and the tissue specimens collected from the left lobe, immediately frozen in liquid nitrogen and stored at −80 °C until being processed for biochemical assays, or cut at cryostat for histochemical assay (tissue sections 8 μm thick, −24 °C; Leica CM 1850 cryostat model, Leica Biosystems Italia, Milano, Italy) [ 64 ].…”

“…Fatty acid profile has been analyzed by a ThermoFisher Scientific DSQII GC/MS system (TraceDSQII mass spectrometer, TraceGCUltra gascromatograph), Xcalibur MS Software Version 2.1 (including NIST Mass Spectral Library (NIST 08) and Wiley Registry of Mass Spectral Data 8th Edition for assignment of chemical structures to chromatographic peaks) [ 64 ]. To proceed to fatty acid assay, 5 µL aliquots of rat liver extracts were dissolved in 1 mL methanolic HCl (2N) into reaction vials.…”