| Grant number: | 11/18746-3 |
| Support Opportunities: | Regular Research Grants |
| Start date: | May 01, 2012 |
| End date: | April 30, 2014 |
| Field of knowledge: | Agronomical Sciences - Veterinary Medicine - Animal Clinics and Surgery |
| Principal Investigator: | Paulo César Ciarlini |
| Grantee: | Paulo César Ciarlini |
| Host Institution: | Faculdade de Medicina Veterinária (FMVA). Universidade Estadual Paulista (UNESP). Campus de Araçatuba. Araçatuba , SP, Brazil |
Abstract
Recently it was demonstrated that chronic renal failure (CRF) causes oxidative stress in cats and dogs. Among the many toxins that concentrate on IRC, p-cresol and guanidines are the most widely investigated in human patients. In this species, these toxins cause uremic neutrophil dysfunction, specifically promotes the inhibition of neutrophil oxidative metabolism, thus affecting their bactericidal function. Our research group has obtained evidence that the in vitro oxidative metabolism of neutrophils from healthy dogs is affected when incubated as uremic serum, urea and creatinine, but so far no studies on the specific effect of uremic toxins p-cresol and guanidine on Neutrophil function in this species. In this sense, this study aims to measure plasma concentrations of p-cresol and guanidine in dogs with CRF and test the hypothesis that the similarity to what occurs in humans the increase of such toxins is associated with alterations of oxidative metabolism and apoptosis of neutrophils. This will standardized the methodology to quantify by chromatography liquid phase plasma concentrations of both toxins in dogs healthy controls and with CRF. It will also be evaluated and compared "in vitro" the specific effect of p-cresol and guanidine on the production of superoxide, lipid peroxidation and the rate of apoptosis of neutrophils isolated from dogs healthy controls, whereas plasma concentrations were observed "in vivo "in dogs with CRF. Cells isolated from 20 healthy dogs will be incubated in media RPMI, with and without p-cresol, guanidine and plasma of dogs with CRF. The oxidative metabolism of neutrophils will be rated by flow cytometry capillary using the probe hidroetidina and by the method cytochemical reduction the nitroblue tetrazolium. Apoptosis and viability of neutrophils is measured in capillary flow cytometry system using Annexin V-PE and by morphometry. Oxidative stress is evaluated by measuring plasma thiobarbituric reactive species, total antioxidant status, uric acid, albumin and total bilirubin. (AU)
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