RESUMEN
Injury due to cold ischemia reperfusion (I/R) is a major cause of primary graft non-function following liver transplantation. We postulated that I/R-induced cellular damage during liver transplantation might affect the secretory pathway, particularly at the endoplasmic reticulum (ER). We examined the involvement of ER stress in organ preservation, and compared cold storage in University of Wisconsin (UW) solution and in Institute Georges Lopez-1 (IGL-1) solution. In one group of rats, livers were preserved in UW solution for 8 h at 4 °C, and then orthotopic liver transplantation was performed according to Kamada's cuff technique. In another group, livers were preserved in IGL-1 solution. The effect of each preservation solution on the induction of ER stress, hepatic injury, mitochondrial damage and cell death was evaluated. As expected, we found increased ER stress after liver transplantation. IGL-1 solution significantly attenuated ER damage by reducing the activation of three pathways of unfolded protein response and their effector molecules caspase-12, C/EBP homologous protein-10, X-box-binding protein 1, tumor necrosis factor-associated factor 2 and eukaryotic translation initiation factor 2. This attenuation of ER stress was associated with a reduction in hepatic injury and cell death. Our results show that IGL-1 solution may be a useful means to circumvent excessive ER stress reactions associated with liver transplantation, and may optimize graft quality.
Asunto(s)
Trasplante de Hígado , Hígado/metabolismo , Soluciones Preservantes de Órganos/farmacología , Preservación de Órganos/métodos , Transducción de Señal/efectos de los fármacos , Adenosina/farmacología , Alopurinol/farmacología , Animales , Apoptosis/efectos de los fármacos , Caspasa 12/genética , Caspasa 12/metabolismo , Isquemia Fría , Frío , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Estrés del Retículo Endoplásmico/efectos de los fármacos , Expresión Génica/efectos de los fármacos , Glutatión/farmacología , Insulina/farmacología , Hígado/patología , Masculino , Rafinosa/farmacología , Ratas , Factores de Transcripción del Factor Regulador X , Daño por Reperfusión/metabolismo , Daño por Reperfusión/patología , Daño por Reperfusión/prevención & control , Transducción de Señal/genética , Factor 2 Asociado a Receptor de TNF/genética , Factor 2 Asociado a Receptor de TNF/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Respuesta de Proteína Desplegada/efectos de los fármacos , Respuesta de Proteína Desplegada/genéticaRESUMEN
The Institut Georges Lopez preservation solution (IGL-1) is a serum-free organ preservative that has been shown to protect steatotic livers against hepatic ischemia-reperfusion injury. Although several hypotheses have been proposed to explain the graft protection mechanisms induced by IGL-1 solution, they have not been fully investigated. This review assessed possible IGL-1 mechanisms responsible for the increased liver tolerance of ischemia-reperfusion injury with special emphasis on vasodilatator mediators such as nitric oxide, on oxidative stress prevention, on protection against mitochondrial damage, and finally on induction of cytoprotective factors.
Asunto(s)
Hígado Graso/complicaciones , Soluciones Preservantes de Órganos , Daño por Reperfusión/prevención & control , Humanos , Daño por Reperfusión/etiologíaRESUMEN
AIM: Static preservation solution is critical for liver graft outcomes, especially when steatosis is present. Institut Georges Lopez (IGL)-1 solution protects fatty livers effectively against cold ischemia reperfusion injury. Its benefits are mediated by nitric oxide and prevention of oxidative stress. The supplementation of IGL-1 with epidermal growth factor (EGF) enhances steatotic graft preservation by increasing adenosine triphosphate content, thereby mitigating oxidative stress and mitochondrial damage. METHODS: After steatotic livers were preserved for 24 hours in IGL-1 solution with or without EGF supplements, they were perfused ex vivo for 2 hours at 37°C. The benefits of EGF were assessed by evidences of hepatic damage and function--transaminases, bile production, and flow rate--as well as by other factors presumably associated with the poor tolerance of fatty livers toward cold ischemia-reperfusion injury (IRI)--energy metabolism, mitochondrial damage, oxidative stress, eNOS activity and proinflammatory interleukin (IL) beta content. RESULTS: Steatotic livers preserved in IGL-1 solutions supplemented with EGF (10 µg/L) showed lower transaminase levels, greater bile production, and ameliorated flow rates when compared to IGL-1 alone. In addition, energy metabolism deterioration, mitochondrial damage, oxidative stress, and cytokine IL-1 beta release were prevented. CONCLUSION: EGF addition to IGL-1 increased fatty liver graft preservation, thereby reducing steatotic liver damage against cold IRI.
Asunto(s)
Factor de Crecimiento Epidérmico/administración & dosificación , Hígado Graso/patología , Preservación Biológica , Animales , Western Blotting , Hígado Graso/metabolismo , Hígado Graso/fisiopatología , Óxido Nítrico Sintasa de Tipo III/metabolismo , Estrés Oxidativo , Ratas , SolucionesRESUMEN
Institute Georges Lopez preservation solution (IGL-1) has been demonstrated to be useful for fatty liver preservation. The mechanisms responsible for this effective graft protection against ischemia-reperfusion injury are pivotal actions on generation of nitric oxide a diffusible molecule with vasodilator properties, that facilitates the up-regulation of other well-known cytoprotective genes, such as hypoxia-inducible factor-1 alpha (HIF-1alpha) and heme-oxygenase 1 (HO-1). During normoxic reperfusion, the presence of nitric oxide permits HIF-1alpha accumulation to inhibit prolyl-hydoxylases, thus promoting an additional overexpression of the HO-1 in steatotic and nonsteatotic graft livers preserved in IGL-1.
Asunto(s)
Hígado Graso/metabolismo , Trasplante de Hígado/patología , Soluciones Preservantes de Órganos , Preservación de Órganos/métodos , Adenosina/efectos adversos , Alopurinol/efectos adversos , Agregación Eritrocitaria/efectos de los fármacos , Hígado Graso/patología , Hígado Graso/cirugía , Glutatión/efectos adversos , Hemo-Oxigenasa 1/genética , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Insulina/efectos adversos , Óxido Nítrico Sintasa/metabolismo , Soluciones Preservantes de Órganos/efectos adversos , Selección de Paciente , Rafinosa/efectos adversos , Regulación hacia ArribaRESUMEN
University of Wisconsin (UW) preservation solution is considered an effective flush and cold storage liquid. However, recent studies have provided evidence of the hyperaggregating effect on human red blood cells (RBC) of hydroxyethyl starch (HES), one of the components of the UW solution. In contrast, preservation solutions containing polyethylene glycol (PEG) have been found to be effective for organ preservation. The aim of this study was to compare the effects of HES (50 g/L); PEG 20 kDa (50 and 30 g/L), and PEG35 kDa (1.05 g/L) added to UW on the rheologic parameters of human RBC at 4 degrees C. Sedimentation rate was measured by the Westergren procedure and blood viscosity evaluated at high shear rates using a cone/plate viscometer. Alterations in RBC morphology and aggregation were evaluated by light microscopy. RBC sedimentation and viscosity were not affected by the inversion of Na+ and K+ concentrations in UW, but were increased by HES. PEGs appeared to reduce RBC deformability with concomitant inhibition of RBC aggregation. These results were consistent with reduced viscosity for PEG-containing solutions. In conclusion, the use of PEG did not change the physiologic function of human RBCs and thus may be an alternative to HES in UW liquids.
Asunto(s)
Viscosidad Sanguínea/efectos de los fármacos , Agregación Eritrocitaria/efectos de los fármacos , Derivados de Hidroxietil Almidón/farmacología , Soluciones Preservantes de Órganos/farmacología , Polietilenglicoles/farmacología , Adenosina/farmacología , Alopurinol/farmacología , Deformación Eritrocítica/efectos de los fármacos , Glutatión/farmacología , Humanos , Insulina/farmacología , Potasio/farmacología , Rafinosa/farmacología , Resistencia al Corte , Sodio/farmacología , Estrés MecánicoRESUMEN
Recent reports argue that the performance of University of Wisconsin (UW) solution is limited by the presence of hydroxyethyl starch (HES) as an additive, since HES could be responsible for human red blood cell aggregation. We investigated the effect on rat liver preservation of replacing HES in UW solution by polyethylene glycols (PEG20 and PEG35) at two concentrations. An isolated perfused rat liver model was used. Six groups of preserved livers (n = 7 for each group) were compared to controls (nonpreserved livers, n = 7). The following preservation solutions were assayed: UW without oncotic supply, UW-HES (0.25 mmol/L), UW-PEG20 (0.03 and 0.25 mmol/L), and UW-PEG35 (0.03 and 0.25 mmol/L). After 24-hour cold storage, the livers were perfused for 120 minutes at 37 degrees C with oxygenated Krebs-Henseleit solution. During perfusion, transaminase release, portal and bile flows, and bromosulfophthalein (BSP) clearance were assessed. Results showed that the omission of oncotic supply in UW statistically increased ALT and AST release in perfusate and decreased bile and portal flows. PEG addition in UW solution, especially PEG35 at 0.25 mmol/L, effectively protected the rat liver graft from the onset of hypothermic ischemia/reperfusion damage. In conclusion, data reported here reveal that oncotic supply is essential for liver preservation and that HES can be effectively replaced by PEG in UW solution.
Asunto(s)
Hígado/fisiología , Soluciones Preservantes de Órganos/farmacología , Polietilenglicoles/farmacología , Adenosina , Alanina Transaminasa/sangre , Alopurinol , Animales , Aspartato Aminotransferasas/sangre , Bilis/metabolismo , Glutatión , Insulina , Hígado/efectos de los fármacos , Pruebas de Función Hepática , Masculino , Sistema Porta/efectos de los fármacos , Sistema Porta/fisiología , Rafinosa , Ratas , Ratas Sprague-DawleyAsunto(s)
Adenosina , Alopurinol , Glutatión , Insulina , Hígado , Soluciones Preservantes de Órganos , Preservación de Órganos/métodos , Glicoles de Propileno/farmacología , Rafinosa , Alanina Transaminasa/sangre , Fosfatasa Alcalina/sangre , Animales , Aspartato Aminotransferasas/sangre , Creatinina/sangre , Hígado/efectos de los fármacos , Hígado/fisiología , Circulación Hepática/fisiología , Pruebas de Función Hepática , Porcinos , Urea/sangreRESUMEN
We used the isolated perfused rat liver model (IPRL) to assess parenchymal and nonparenchymal cell integrity after different conditions of storage and reperfusion. Two studies were performed. In study 1, the IPRL was applied to evaluate the effects of 30 min of normothermic reperfusion with Elohes solution, enriched William's medium (Wif), or Carolina rinse solution (CRS) following 24 h of cold preservation in high-K+ or high-Na+ UW solution. As indicated by creatine kinase-BB (CK-BB) release, reperfusion with CRS provided greater protection of endothelial cells after storage in high-K+ UW solution than after storage in high-Na+ UW solution. In study 2, livers were cold-preserved (24 h, 4 degrees C) in either high-K+ or high-Na+ UW solution, then flushed with either CRS or Wif solution at room temperature before reperfusion (120 min, 37 degrees C) with 5% albumin-William's medium E. There was no statistical difference between the rinse solutions for bile flow and transaminases release. However, CRS improved bile indocyanine green excretion, which is known to be a marker of parenchymal and nonparenchymal cell integrity. Therefore, we can assume that this rinse solution protects rat liver grafts from reperfusion-induced microvascular damage.
Asunto(s)
Criopreservación , Hígado/patología , Preservación de Órganos , Daño por Reperfusión/patología , Animales , Creatina Quinasa/metabolismo , Endotelio Vascular/enzimología , Endotelio Vascular/patología , Isoenzimas , Hígado/efectos de los fármacos , Hígado/enzimología , Pruebas de Función Hepática , Masculino , Soluciones Preservantes de Órganos/farmacología , Ratas , Ratas Wistar , Daño por Reperfusión/enzimologíaRESUMEN
The isolated perfused rat liver model was used to assess graft viability after 24 h of cold preservation. Two solutions were compared for liver preservation: Belzer's original UW solution (high-K+ UW) and a solution containing the same components but with inverted concentrations of sodium and potassium (high-Na+ UW). During the 120 min of normothermic reperfusion, livers preserved in the high-Na+ UW solution released lower levels of creatine kinase-BB isoenzyme, transaminases (ALT and AST), and potassium than those preserved in the high-K+ UW solution. Bile flow and biliary excretion of indocyanine green increased when livers were preserved in the high-Na+ UW solution. We found no statistical differences for oxygen consumption and tissue ATP concentration. The results of this study support the concept that a high-Na+ UW solution is a more effective means of preserving rat livers, at least after 24 h of cold-storage and 120 min of reperfusion in the isolated perfused model, than the original high-K+ UW solution. Liver preservation in the high-Na+ UW solution reduces damage to sinusoidal endothelial and hepatocellular cells. The use of an extracellular-like Belzer cold storage solution eliminates potassium-related problems in cold preservation and subsequent normothermic reperfusion while keeping all the qualities of the original UW solution.