Background/aim. Sickle cell disease is a worldwide-distributed autosomal recessive genetic red cell disorder. Hepatic dysfunction and liver damage is present in sickle cell disease, but its pathogenesis is only partially known. Design and Methods. Transgenic mouse model for sickle cell disease (SAD mice) and wild-type mice were exposed to ischemic/reperfusion protocol. The following parameters were evaluated: hematological profile, transaminases and bilirubin levels, liver pathology, mRNA levels of nuclear factor-kB p65, endothelial NO synthase, inducible NO synthase, heme oxygenase-1 and phosphodiesterase-1-2-3-4 genes in hepatocytes obtained by laser-capture-microdissection. Protein expression of nuclear factor-kB p65 and phospho- nuclear factor-kB p65, heme oxygenase-1, biliverdin reductase, heat-shock-protein-70,-27 and peroxiredoxin-6 were analysed by immunoblot. A subgroup of SAD mice was treated with the phosphodiesterase-4 inhibitor Rolipram (30 mg/Kg/day by gavage) during the ischemic/reperfusion stress. Results. In SAD mice the ischemic/reperfusion stress induced liver damage compatible with sickle cell disease hepatopathy, which was associated with (i) lack of hypoxia induced nuclear factor-kB p65 activation; (ii) unbalance in endothelial /inducible NO synthase response to ischemic/reperfusion stress; (iii) lack of hypoxia induced heme oxygenase-1/ biliverdin reductase increased expression paralleled by the compensatory increased in heat-shock-protein-70,-27 and peroxiredoxin-6 expression; (iv) up-regulation of the phosphodiesterase-1,-2,-3,-4 genes. In SAD mice the phosphodiesterase-4 inhibitor Rolipram attenuates the ischemic/reperfusion related microcirculatory dysfunction, reduces the inflammatory cell infiltration and induced the heme oxygenase-1/ biliverdin reductase cytoprotective systems. Conclusions. In SAD mice, sickle cell hepatopathy is associated with perturbation of nuclear factor-kB p65 signalling with imbalance of endothelial /inducible nitric oxide synthase levels, lack of heme oxygenase-1/ biliverdin reductase expression and up-regulation of two novel cytoprotective systems: heat-shock-protein-27 and peroxiredoxin-6.
Abnormal modulation of cell protective systems in response to ischemic/reperfusion injury is important in the development of mouse sickle cell hepatopathy
SICILIANO, Angela;MALPELI, Giorgio;SCARPA, Aldo;OLIVIERI, Oliviero;AMATO, Eliana;CORROCHER, Roberto;DE FRANCESCHI, Lucia
2011-01-01
Abstract
Background/aim. Sickle cell disease is a worldwide-distributed autosomal recessive genetic red cell disorder. Hepatic dysfunction and liver damage is present in sickle cell disease, but its pathogenesis is only partially known. Design and Methods. Transgenic mouse model for sickle cell disease (SAD mice) and wild-type mice were exposed to ischemic/reperfusion protocol. The following parameters were evaluated: hematological profile, transaminases and bilirubin levels, liver pathology, mRNA levels of nuclear factor-kB p65, endothelial NO synthase, inducible NO synthase, heme oxygenase-1 and phosphodiesterase-1-2-3-4 genes in hepatocytes obtained by laser-capture-microdissection. Protein expression of nuclear factor-kB p65 and phospho- nuclear factor-kB p65, heme oxygenase-1, biliverdin reductase, heat-shock-protein-70,-27 and peroxiredoxin-6 were analysed by immunoblot. A subgroup of SAD mice was treated with the phosphodiesterase-4 inhibitor Rolipram (30 mg/Kg/day by gavage) during the ischemic/reperfusion stress. Results. In SAD mice the ischemic/reperfusion stress induced liver damage compatible with sickle cell disease hepatopathy, which was associated with (i) lack of hypoxia induced nuclear factor-kB p65 activation; (ii) unbalance in endothelial /inducible NO synthase response to ischemic/reperfusion stress; (iii) lack of hypoxia induced heme oxygenase-1/ biliverdin reductase increased expression paralleled by the compensatory increased in heat-shock-protein-70,-27 and peroxiredoxin-6 expression; (iv) up-regulation of the phosphodiesterase-1,-2,-3,-4 genes. In SAD mice the phosphodiesterase-4 inhibitor Rolipram attenuates the ischemic/reperfusion related microcirculatory dysfunction, reduces the inflammatory cell infiltration and induced the heme oxygenase-1/ biliverdin reductase cytoprotective systems. Conclusions. In SAD mice, sickle cell hepatopathy is associated with perturbation of nuclear factor-kB p65 signalling with imbalance of endothelial /inducible nitric oxide synthase levels, lack of heme oxygenase-1/ biliverdin reductase expression and up-regulation of two novel cytoprotective systems: heat-shock-protein-27 and peroxiredoxin-6.
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