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Nephrology Dialysis Transplantation, Vol 12, Issue 1 174-179, Copyright © 1997 by Oxford University Press

ORIGINAL ARTICLES

A simple method to identify NBT-positive cells in isolated glomeruli

M Romero, J Mosquera and B Rodriguez-Iturbe
Instituto de Investigaciones Clinicas, Universidad del Zulia, Venezuela; Renal Service, Hospital Universitario and Instituto de Investigaciones Biomedicas (INBIOMED), Maracaibo, Venezuela; Corresponding author at: Unidad de Dialisis y Trasplante Renal, Hospital Universitario (9 piso), Aparado Postal 1430, Maracaibo, 4001-A, Venezuela

Background. Reactive oxygen radicals are probably involved in the pathogenesis of human and experimental models of renal disease, yet current methods are inadequate to quantify and identify the cells producing reactive oxygen radicals. Methods and Results. We used the nitroblue tetrazolium reaction to determine superoxide anion production in glomerular cells in phorbol myristate-stimulated glomerular suspensions and in isolated glomeruli from rats with nephrotoxic nephritis, ureteral obstruction, and puromycin aminonucleoside nephrosis. We were also able to identify these nitroblue tetrazolium+cells using specific appropriate antibodies. When the technique was tested in conditions known to increase reactive oxygen radicals, as phorbol myristate-stimulated glomeruli and glomeruli from animals with nephrotoxic nephritis and ureteral obstruction, increased number of nitroblue tetrazolium+cells were found. These cells were identified as glomerular intrinsic cells (Thy-1 +) or infiltrating leukocytes (leukocyte common antigen+ or antineutrophil+). Conclusions. This method may be useful to determine cells participating in glomerular damage induced by reactive oxygen radicals. Keywords: glomerulus; nitroblue tetrazolium; reactive oxygen radicals
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