title: Sirtuin 1 in rat orthotopic liver transplantation: an IGL-1 preservation solution approach.
abstract: OBJECTIVE
  To investigate the possible involvement of Sirtuin 1 (SIRT1) in rat orthotopic liver transplantation (OLT), when Institute Georges Lopez 1 (IGL-1) preservation solution is enriched with trimetazidine (TMZ).
  
  
  METHODS
  Male Sprague-Dawley rats were used as donors and recipients. Livers were stored in IGL-1 preservation solution for 8h at 4 °C, and then underwent OLT according to Kamada's cuff technique without arterialization. In another group, livers were stored in IGL-1 preservation solution supplemented with TMZ, at 10(-6) mol/L, for 8 h at 4 °C and then underwent OLT. Rats were sacrificed 24 h after reperfusion, and liver and plasma samples were collected. Liver injury (transaminase levels), mitochondrial damage (glutamate dehydrogenase activity) oxidative stress (malondialdehyde levels), and nicotinamide adenine dinucleotide (NAD(+)), the co-factor necessary for SIRT1 activity, were determined by biochemical methods. SIRT1 and its substrates (ac-FoxO1, ac-p53), the precursor of NAD(+), nicotinamide phosphoribosyltransferase (NAMPT), as well as the phosphorylation of adenosine monophosphate activated protein kinase (AMPK), p-mTOR, p-p70S6K (direct substrate of mTOR), autophagy parameters (beclin-1, LC3B) and MAP kinases (p-p38 and p-ERK) were determined by Western blot.
  
  
  RESULTS
  Liver grafts preserved in IGL-1 solution enriched with TMZ presented reduced liver injury and mitochondrial damage compared with those preserved in IGL-1 solution alone. In addition, livers preserved in IGL-1 + TMZ presented reduced levels of oxidative stress. This was consistent with enhanced SIRT1 protein expression and elevated SIRT1 activity, as indicated by decreased acetylation of p53 and FoxO1. The elevated SIRT1 activity in presence of TMZ can be attributed to the enhanced NAMPT protein and NAD(+)/NADH levels. Up-regulation of SIRT1 was consistent with activation of AMPK and inhibition of phosphorylation of mTOR and its direct substrate (p-p70S6K). As a consequence, autophagy mediators (beclin-1 and LC3B) were over-expressed. Furthermore, MAP kinases were regulated in livers preserved with IGL-1 + TMZ, as they were characterized by enhanced p-ERK and decreased p-p38 protein expression.
  
  
  CONCLUSIONS
  Our study shows that IGL-1 preservation solution enriched with TMZ protects liver grafts from the IRI associated with OLT, through SIRT1 up-regulation.
authors: Eirini Pantazi, Mohamed Amine Zaouali, Mohamed Bejaoui, Emma Folch-Puy, Joan Roselló-Catafau, Experimental Hepatic Ischemia, Reperfusion Unit, Institute of Biomedical Research of Barcelona, Barcelona, 08036 Catalonia, Spain.
authors: Eirini
authors: E
authors: Pantazi
authors: Eirini Pantazi, Mohamed Amine Zaouali, Mohamed Bejaoui, Emma Folch-Puy, Joan Roselló-Catafau, Experimental Hepatic Ischemia, Reperfusion Unit, Institute of Biomedical Research of Barcelona, Barcelona, 08036 Catalonia, Spain.
authors: Mohamed Amine
authors: MA
authors: Zaouali
authors: Eirini Pantazi, Mohamed Amine Zaouali, Mohamed Bejaoui, Emma Folch-Puy, Joan Roselló-Catafau, Experimental Hepatic Ischemia, Reperfusion Unit, Institute of Biomedical Research of Barcelona, Barcelona, 08036 Catalonia, Spain.
authors: Mohamed
authors: M
authors: Bejaoui
authors: Eirini Pantazi, Mohamed Amine Zaouali, Mohamed Bejaoui, Emma Folch-Puy, Joan Roselló-Catafau, Experimental Hepatic Ischemia, Reperfusion Unit, Institute of Biomedical Research of Barcelona, Barcelona, 08036 Catalonia, Spain.
authors: Emma
authors: E
authors: Folch-Puy
authors: Eirini Pantazi, Mohamed Amine Zaouali, Mohamed Bejaoui, Emma Folch-Puy, Joan Roselló-Catafau, Experimental Hepatic Ischemia, Reperfusion Unit, Institute of Biomedical Research of Barcelona, Barcelona, 08036 Catalonia, Spain.
authors: Hassen
authors: H
authors: Ben Abdennebi
authors: Eirini Pantazi, Mohamed Amine Zaouali, Mohamed Bejaoui, Emma Folch-Puy, Joan Roselló-Catafau, Experimental Hepatic Ischemia, Reperfusion Unit, Institute of Biomedical Research of Barcelona, Barcelona, 08036 Catalonia, Spain.
authors: Ana Teresa
authors: AT
authors: Varela
authors: Eirini Pantazi, Mohamed Amine Zaouali, Mohamed Bejaoui, Emma Folch-Puy, Joan Roselló-Catafau, Experimental Hepatic Ischemia, Reperfusion Unit, Institute of Biomedical Research of Barcelona, Barcelona, 08036 Catalonia, Spain.
authors: Anabela Pinto
authors: AP
authors: Rolo
authors: Eirini Pantazi, Mohamed Amine Zaouali, Mohamed Bejaoui, Emma Folch-Puy, Joan Roselló-Catafau, Experimental Hepatic Ischemia, Reperfusion Unit, Institute of Biomedical Research of Barcelona, Barcelona, 08036 Catalonia, Spain.
authors: Carlos Marques
authors: CM
authors: Palmeira
authors: Eirini Pantazi, Mohamed Amine Zaouali, Mohamed Bejaoui, Emma Folch-Puy, Joan Roselló-Catafau, Experimental Hepatic Ischemia, Reperfusion Unit, Institute of Biomedical Research of Barcelona, Barcelona, 08036 Catalonia, Spain.
authors: Joan
authors: J
authors: Roselló-Catafau
mesh_terms: D000818:Animals; D001343:Autophagy / Q000187:drug effects; D015415:Biomarkers / Q000097:blood; D050377:Cold Ischemia; D006085:Graft Survival / Q000187:drug effects; D008099:Liver / Q000187:drug effects* / Q000201:enzymology / Q000601:surgery*; D016031:Liver Transplantation / Q000009:adverse effects / Q000379:methods*; D008297:Male; D008930:Mitochondria, Liver / Q000187:drug effects / Q000201:enzymology; D009926:Organ Preservation / Q000009:adverse effects / Q000379:methods*; D019188:Organ Preservation Solutions / Q000494:pharmacology*; D018384:Oxidative Stress / Q000187:drug effects; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000097:blood / Q000201:enzymology / Q000209:etiology / Q000517:prevention & control*; D015398:Signal Transduction / Q000187:drug effects; D056564:Sirtuin 1 / Q000378:metabolism*; D013997:Time Factors; D014292:Trimetazidine / Q000494:pharmacology; D015854:Up-Regulation
chemical_list: D015415:Biomarkers; C504757:IGL-1 solution; D019188:Organ Preservation Solutions; C488368:Sirt1 protein, rat; D056564:Sirtuin 1; D014292:Trimetazidine
keywords: IGL-1 preservation solution; Ischemia-reperfusion injury; Liver transplantation; Sirtuin 1; Trimetazidine