Post-exercise proteinuria is a common phenomenon in healthy subjects. Previous studies have used albumin (Alb) and beta 2-microglobulin (beta 2-m) molecules as representatives of high- and low-molecular-weight proteins. Recently, more specific markers of the human kidney proximal tubule have been used to identify the precise site of alterations. Active male subjects underwent two strenuous runs, one 400-m run and one 3000-m run. Urine was collected from the subjects before and after each event. Total protein (TP), Alb, alpha 1-microglobulin (alpha 1-m), beta 2-m, intestinal alkaline phosphatase (IAP), tissue-nonspecific alkaline phosphatase (TNAP) and N-acetyl-beta-D-glucosaminidase (NAG) were determined for each sample. The short-distance run (400 m) resulted in the largest increases (P < or = 0.05) in TP (31-fold), Alb (100-fold) and beta 2-m (164-fold) as compared to the long-distance run (3000-m). The alpha 1-m excretion rates were increased to a lesser extent by the exercises. The IAP activity was slightly increased (+90%) by the 400-m run while the TNAP and NAG activities showed a 6.8-fold and a 3.6-fold increase, respectively, after this event. Smaller increases were recorded for the long-distance run (P = 0.05). To conclude, the present investigation showed that: (1) post-exercise proteinuria is related to the absolute intensity of exercise; (2) the impairment of protein reabsorption is revealed better by changes in Alb and beta 2-m; (3) changes in TNAP and NAG activities could reveal biochemical modifications that occur in the proximal tubule, particularly at the S1-S2 segment.