RESUMO
We report the molecular cloning of the cDNA sequence for pig peripheral benzodiazepine receptor (PBR) by using RT-PCR and 5'/3' terminal extension. Three different transcripts (long, middle, and short) are identified. The open reading frame (ORF) of the longest PBR mRNA encodes a deduced polypeptide of 169 amino acids with a calculated molecular weight of 18,609 Da and an estimated pI of 9.70, which corresponds to the authentic PBR of other mammalian species. The middle transcript (PBR-M) contains a 141-codon ORF, which is consistent with that of the authentic PBR, but lacks a region of 84 bp so that its encoded polypeptide lacks a region of 28 amino acids from 35 to 62 of the authentic PBR polypeptide. The short transcript (PBR-S) contains a 104-codon ORF, which overlaps that of the authentic PBR, but lacks a region of 211 bp so that its encoded polypeptide lacks a region of 65 amino acids of the N-terminal of the authentic PBR. The pig PBR gene was mapped to the telomeric end of SSC5p. In addition, PBR mRNA was the more abundant detected form in pig tissues and in warm kidney that underwent ischemia suggesting functional implications of PBR during the renal repair process.
Assuntos
Processamento Alternativo/genética , Mapeamento Cromossômico , Clonagem Molecular , Receptores de GABA/genética , Análise de Sequência de DNA , Suínos/genética , Glândulas Suprarrenais/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Mapeamento Cromossômico/métodos , Clonagem Molecular/métodos , DNA Complementar/isolamento & purificação , Feminino , Rim/metabolismo , Masculino , Mitocôndrias/metabolismo , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/métodos , Isoformas de Proteínas , RNA Mensageiro/análise , Receptores de GABA/metabolismo , Reperfusão/efeitos adversos , Traumatismo por Reperfusão/metabolismo , Análise de Sequência de DNA/métodos , Distribuição Tecidual , Isquemia Quente/efeitos adversosRESUMO
In organ transplantation, ischemia-reperfusion injury (IRI) has been implicated in delayed graft function (DGF) as well as in short- and long-term complications. Using an autotransplant pig kidney model, changes in renal function and morphology were determined after different periods of cold ischemia in kidneys preserved in the University of Wisconsin solution (UW), high-Na(+) version of UW (HEH) or Celsior (CEL) a newly developed high-Na(+) solution, with or without trimetazidine (TMZ). Kidney function was better preserved in CEL, UW and particularly HEH in combination with TMZ, particularly after 48 and 72 h. Mitochondria integrity was improved in TMZ-preserved groups. This study indicates that TMZ is efficiently protective against IRI even after prolonged preservation and in different preservation solutions.
Assuntos
Criopreservação , Mitocôndrias/efeitos dos fármacos , Traumatismo por Reperfusão/tratamento farmacológico , Trimetazidina/farmacologia , Adenosina/farmacologia , Alopurinol/farmacologia , Animais , Biomarcadores , Dissacarídeos/farmacologia , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Eletrólitos/farmacologia , Glutamatos/farmacologia , Glutationa/farmacologia , Histidina/farmacologia , Insulina/farmacologia , Testes de Função Renal/métodos , Transplante de Rim/métodos , Transplante de Rim/mortalidade , Manitol/farmacologia , Mitocôndrias/fisiologia , Mitocôndrias/ultraestrutura , Preservação de Órgãos/métodos , Soluções para Preservação de Órgãos/farmacologia , Rafinose/farmacologia , Receptores de GABA-A/efeitos dos fármacos , Receptores de GABA-A/fisiologia , Traumatismo por Reperfusão/prevenção & controle , Suínos , Fatores de Tempo , Transplante Autólogo/métodos , Transplante Autólogo/mortalidadeRESUMO
Ischemia-reperfusion injury (IRI) is associated with an increased risk of acute rejection, delayed graft function, or chronic graft dysfunction. Mitochondria plays a central role in this process. Using an autotransplant pig kidney model, changes in renal function and morphology were determined after different periods of cold ischemia in kidneys preserved in the University of Wisconsin solution (UW), high-Na(+) version of UW (HEH) or Celsior (CEL) a newly developed high-Na(+) solution, with or without trimetazidine (TMZ). Kidney function was better preserved in HEH after 24 hr and particularly 48- and 72-hr cold storage than in CEL and UW. TMZ improved the preservation quality when added to the different solutions tested, particularly after 48- and 72-hr cold storage. Interstitial fibrosis and tubular atrophy were reduced in HEH with TMZ. CD4(+) T-cell infiltration was also modulated by the preservation conditions. Peripheral-type benzodiazepine receptor (PBR) positive cells infiltration was also modulated by preservation conditions. TMZ was efficient to reduce IRI when added in the various preservation solutions. These results suggest that protection of the mitochondrial function should be a major target to limit IRI. In addition, this study outlines the role of CD4(+) T cells and PBR expression in inflammatory responses after IRI.