RT Journal Article
T1 Glutamyl aminopeptidase in microvesicular and exosomal fractions of urine is related with renal dysfunction in cisplatin-treated rats
A1 Quesada, Andrés
A1 Segarra, Ana Belén
A1 Montoro-Molina, Sebastián
A1 de Gracia, María Del Carmen
A1 Osuna, Antonio
A1 O'Valle, Francisco
A1 Gómez-Guzmán, Manuel
A1 Vargas, Félix
A1 Wangensteen, Rosemary
K1 Animales
K1 Antígenos CD13
K1 Peso corporal
K1 Centrifugación
K1 Cisplatino
K1 Creatinina
K1 Técnicas para inmunoenzimas
K1 Espacio extracelular
K1 Glutamil aminopeptidasa
K1 Pruebas de función renal
K1 Neoplasias
K1 Ratas
K1 Aumento de peso
AB PURPOSE:The aim of this work was to investigate if the content of glutamyl aminopeptidase (GluAp) in microvesicular and exosomal fractions of urine is related with renal dysfunction in cisplatin-treated rats.METHODS:Urine samples were collected 24 hours after injection of cisplatin (7 mg/kg, n = 10) or saline serum (n = 10), and they were subjected to differential centrifugation at 1.000, 17.000 and 200.000 g to obtain microvesicular and exosomal fractions. GluAp was measured with a commercial ELISA kit in both fractions. Serum creatinine (SCr) and body weight were measured 15 days after treatment. We analyzed if early excretion of GluAp in microsomal and exosomal fractions was correlated with final SCr and body weight increase. In a second experiment, enzymatic activities of GluAp and alanyl aminopeptidase (AlaAp) in urine, microvesicular and exosomal fractions were measured three days after injection. We analyzed the correlation of both markers with SCr determined at this point. Finally, we studied the expression of GluAp and extracellular vesicles markers Alix and tumor susceptibility gene (TSG101) in both fractions by immunoblotting.RESULTS:GluAp excretion was increased in all fractions of urine after cisplatin treatment, even if data were normalized per mg of creatinine, per body weight or per total protein content of each fraction. We found significant predictive correlations with SCr concentration, and inverse correlations with body weight increase determined 15 days later. Three days after injection, aminopeptidasic activities were markedly increased in all fractions of urine in cisplatin-treated rats. The highest correlation coefficient with SCr was found for GluAp in microvesicular fraction. Increase of GluAp in microvesicular and exosomal fractions from cisplatin-treated rats was confirmed by immunoblotting. Alix and TSG101 showed different patterns of expression in each fraction.CONCLUSIONS:Determination of GluAp content or its enzymatic activity in microvesicular and exosomal fractions of urine is an early and predictive biomarker of renal dysfunction in cisplatin-induced nephrotoxicity. Measurement of GluAp in these fractions can serve to detect proximal tubular damage independently of glomerular filtration status.
PB Public Library of Science
YR 2017
FD 2017-04-11
LK http://hdl.handle.net/10668/2689
UL http://hdl.handle.net/10668/2689
LA en
NO Quesada A, Segarra AB, Montoro-Molina S, de Gracia MD, Osuna A, O'Valle F, et al. Glutamyl aminopeptidase in microvesicular and exosomal fractions of urine is related with renal dysfunction in cisplatin-treated rats.PLoS One. 2017;12(4):e0175462.
NO This study was supported by the grants PI13/02743 (AO) and PI13/02384 (RW) from the Carlos III Health Institute of Spain, and the Red de Investigación Renal REDinREN RD16/0009/0033 (AO).“FEDER una manera de hacer Europa”.
DS RISalud
RD Apr 12, 2025