Renal prothrombin mRNA is significantly decreased in a hyperoxaluric rat model of nephrolithiasis

Although urinary prothrombin fragment 1 (UPTF1) possesses several hallmarks expected of a regulatory protein in urolithiasis, its precise role remains unknown. To determine the relationship between renal prothrombin (PT), the parent molecule of UPTF1, and lithogenesis, this study quantified and compared levels of renal PT mRNA in healthy rats (n = 10) and rats rendered lithogenic (n = 10) by ingestion of 0.75% ethylene glycol for 8 weeks. Studies included morphological and histological examination of the kidneys with scanning electron microscopy of the urinary filtrates of control and experimental animals. Haematuria and calcium oxalate (CaOx) crystals occurred in the urine of all experimental rats, but not in those of controls. Histological examination showed birefringent nephroliths and associated damage in kidneys of lithogenic rats, which were not seen in the control group. The amounts of total RNA extracted from both groups of rats were similar, but the median ratio of PT to beta-actin transcript of 11.14 x 10(-4) (10.65 x 10(-4) +/- 2.24 x 10(-4)) in the control rats was significantly (p < or = 0.001) reduced to 6.47 x 10(-4) (6.57 x 10(-4) +/- 2.72 x 10(-4)) in the lithogenic group. These results demonstrate that renal PT mRNA is reduced by approximately 42% in lithogenic rats and confirm the existence of a direct association between renal PT synthesis and calculogenesis. Attempts to compare renal PT and urinary levels of PTF1 were unsuccessful because of interference from hepatic PT circulating in the blood, haematuria, and the presence of urinary CaOx crystals. This is the first report of a significant reduction in the renal expression of a urinary protein well documented to inhibit CaOx crystal growth and aggregation in undiluted human urine in vitro.