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Renal enzyme activities in experimental ochratoxin A-induced porcine nephropathy: diagnostic potential of phosphoenolpyruvate carboxykinase and gamma-glutamyl transpeptidase activity.

Enzyme activities have been measured in needle biopsies from kidneys of pigs fed 1 ppm or 0.2 ppm of ochratoxin A for 1-5 wks. After feeding 1 ppm toxin for 1 wk, the activity of cytosolic phosphoenolpyruvate carboxykinase (PEPCK) was decreased by 40% and remained inhibited until the termination of the experiment (5 wk). The activity of gamma-glutamyl transpeptidase, a brush-border enzyme found in the proximal tubules, was reduced to a similar degree and remained inhibited. The activities of hexokinase, a cytosolic enzyme of general distribution in the nephron, and phosphate-dependent glutaminase, a distal tubule enzyme, were not affected. The biopsy results were confirmed by measurements in renal slices taken at the termination of the experiment, except that biopsy samples showed more variation in enzyme activity and a lower PEPCK activity. A guinea pig antibody against the cytosolic form of PEPCK was used to demonstrate that the mitochondrial form of the enzyme, which accounts for a considerable part of the total cellular activity, was not affected by ochratoxin A. When mitochondrial PEPCK activity present in the cytosolic fraction was accounted for, ochratoxin A was found to reduce PEPCK activity by 70-80%. The increase of ochratoxin A exposure from zero through 0.2 ppm to 1 ppm, which resulted in dose-dependent activity decrease of PEPCK and gamma-glutamyl transpeptidase, was accompanied by dose-dependent decrease of renal function, as measured by a reduction of maximal tubular excretion of para-aminohippurate per clearance of inulin (TmPAH/CIn) and an increase in glucose excretion. This suggest that these enzymes are sensitive indicators of ochratoxin A-induced porcine nephropathy. Assuming that porcine nephropathy represents a valid model of endemic (Balkan) nephropathy in humans, the measurement of cytosolic PEPCK and gamma-glutamyl transpeptidase activity in the kidney could be a sensitive test for ochratoxin A-induced disease in humans.