RESUMO
Endostatin, a carboxyl-terminal fragment of collagen XVIII is known as an anti-angiogenic agent, that specifically inhibits the proliferation of endothelial cell and the growth of several primary tumor. We report here the purification and characterization of the recombinant murine endostatin (rmEndostatin) which was expressed in a prokaryotic expression system. This rmEndostatin has similar physiochemical properties of yeast-produced recombinant endostatin, and it also specifically inhibits the proliferation and migration of bovine capillary endothelial cells stimulated by basic fibroblast growth factor. The biological activity of rmEndostatin was also shown by its anti-angiogenic ability on the chorioallantoic membrane of chick embryo in vivo. In this article, we demonstrate the refolding and purification of rmEndostatin, expressed using E. coli system, to a biologically active and soluble form. In addition, these results confirm the activity of endostatin as a potent anti-angiogenic agent.
Assuntos
Inibidores da Angiogênese/genética , Inibidores da Angiogênese/farmacologia , Colágeno/genética , Colágeno/farmacologia , Escherichia coli/genética , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/farmacologia , Inibidores da Angiogênese/isolamento & purificação , Animais , Western Blotting , Bovinos , Movimento Celular/efeitos dos fármacos , Embrião de Galinha , Córion/efeitos dos fármacos , Córion/patologia , Dicroísmo Circular , Colágeno/isolamento & purificação , Colágeno Tipo XVIII , Eletroforese em Gel de Poliacrilamida , Endostatinas , Endotélio Vascular/citologia , Endotélio Vascular/efeitos dos fármacos , Fator 2 de Crescimento de Fibroblastos/farmacologia , Camundongos , Neovascularização Fisiológica/efeitos dos fármacos , Fragmentos de Peptídeos/isolamento & purificação , Dobramento de Proteína , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/farmacologia , Solubilidade , Leveduras/genéticaRESUMO
INTRODUCTION: The aim of this study was to analyze risk factors for delayed graft function (DGF) after deceased donor kidney transplantation and to compare the clinical outcomes of non-DGF versus DGF recipients. PATIENTS AND METHODS: From January 2004 to June 2008, 75/154 kidneys were transplanted into 74 recipients. We classified the recipients into two groups: group 1 (n=61) without DGF and group 2 (n=13) with DGF. RESULTS: On univariate analysis, recipient age (P=.048) cause of brain death (traumatic brain injury vs disease, P=.016), blood urea nitrogen (P=.002), serum creatinine (P=.001), arterial pH (P=.019), and serum sodium level (P=.012) just before organ procurement showed significant differences. On multivariate analysis, the cause of brain death (P=.015, hazard ratio [HR]: 7.086), the terminal serum creatinine>or=1.5 mg/dL before organ procurement (P=.007, HR: 10.132), and recipient age over >or=50 years (P=.021, HR: 7.767) were independent risk factors for the development of DGF. Graft failures occurred among 5/74 recipients with 5-year graft survivals between group 1 and group 2 of 91.7% and 84.6%, respectively. Patient death occurred in five cases, most by due to infection. The 5-year patient survival between groups 1 and 2 were 93.9% and 84.6%, respectively (P = .106). CONCLUSION: The independent risk factors for DGF were the cause of brain death, the terminal creatinine level, and the recipient age. In deceased donor kidney transplantation, DGF may have less effect on long-term patient and graft survivals.