Expression of proteins that inhibit calcium oxalate crystallization in vitro in the urine of normal and stone-forming individuals

Abstract

The factors precipitating clinically active calcium oxalate (CaOx) urolithiasis are not known. This study examined the relationships between urinary proteins that inhibit CaOx crystallization in vitro and the incidence of CaOx urolithiasis. The first hypothesis is that levels of urinary CaOx crystallization inhibitors differ between clinically active stone formers (SFs) and normal individuals. The second hypothesis is that lower levels of urinary CaOx crystallization inhibitors contribute to the two- to threefold greater incidence of CaOx urolithiasis in males compared with females. These hypotheses were derived from previous observations on the expression of urinary inter-alpha-trypsin inhibitor trimer (IalphaTI-trimer) in normal and stone-forming individuals. The proteins of void urine samples from normal volunteers (24 males, 19 females) and CaOx-SFs (26 males, 16 females) were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Immunoreactive IalphaTI-trimer, osteopontin, and prothrombin were detected by immunoblot plus enhanced chemiluminescence; the relative densities of the bands were then determined. With the exception of IalphaTI-trimer (P: </= 0.026, approximately twofold), there was no difference in the relative densities of CaOx crystallization inhibitors in the urine of normal and CaOx stone-forming individuals. Thus, there does not appear to be a generalized increase or decrease in levels of CaOx crystallization inhibitory proteins between normal and CaOx stone-forming individuals. The relative density of IalphaTI-trimer was approximately threefold greater in females than in males (P: </= 0.001). Differences in the relative densities of the other CaOx crystallization inhibitors were small and of questionable physiological importance. These data do not support the hypothesis that males have a greater incidence of CaOx urolithiasis because of a generalized decrease in urinary CaOx crystallization inhibitory protein levels.