Aldosterone metabolites in spontaneously hypertensive rats

Abstract

Several aldosterone metabolites are now known to possess some mineralocorticoid activities. In order to test the hypothesis that these metabolites could contribute to the pathogenesis of hypertension, we studied the aldosterone metabolism in SHR in vitro and in vivo. In vitro experiment, male SHR and WKY rats of 4 and 15 weeks of age were used. The microsome, cytosol and heavy mitochondria fractions from liver and kidney were isolated by ultracentrifuge. 10mg protein/ml of each subcellular fraction was incubated with 3H-aldosterone in Tris-HCl buffer at pH 7.4 containing NADPH, glucose-6-phosphate (G-6-P) and G-6-P dehydrogenase as described by Morris, D.J. et al. (Hypertension, 5 (suppl. I]: I-35-I-40, 1983.). Aldosterone and its metabolites synthesized were extracted with Sep-pak C18 cartridges and separated by HPLC on a reverse phase column. In vivo experiments, the urine of male SHR and WKY rats of 15 weeks old injected 10 microCi 3H-aldosterone intraperitoneally was collected for 48 hours, extracted and analyzed by HPLC. Peaks of steroids from SHR were compared with those from WKY. Incubation of aldosterone with liver microsomes yielded at least 10 polar and 3 less polar metabolites (A-ring reduced metabolites). SHR liver microsomes synthesized larger amounts of 3 polar metabolites than WKY liver microsomes. Liver cytosol, liver heavy mitochondria and kidney subcellular fractions mainly synthesized less polar metabolites, but failed to synthesize as much polar metabolites as liver microsomes. Kidney microsomes and cytosol from 4 weeks old SHR synthesized larger amounts of less polar metabolites compared to those from WKY. In vivo experiment, SHR of 15 weeks of age excreted larger amounts of 2 polar metabolites than WKY. The present study suggests that the difference of metabolism of aldosterone between SHR and WKY observed from an early stage in the liver and the target organ, kidney, may be associated with hypertension or its causative factors, and confirms that aldosterone will be metabolized to several polar and less polar forms by rat liver and kidney subcellular fractions.