Physicochemical characterization of the urinary lipid from humans with nephrotic syndrome

The aim of this study was to investigate the nature of urinary lipid in humans with nephrotic syndrome. Fresh urine specimens were fractionated by centrifugation into a lower cellular fraction and an upper noncellular fraction. Of the 11 urine specimens examined, six cellular fractions contained cells (oval fat bodies) that were laden with anisotropic (birefringent) droplets when viewed by polarizing microscopy. The mean total cholesterol excretions for the five urine specimens without anisotropic droplets and the six urine specimens with anisotropic droplets were 8.7 mg/L and 35.5 mg/L, respectively. The cellular fractions of the six urine specimens with anisotropic droplets are enriched in cholesterol esters relative to whole urine. Upon heating, the anisotropic droplets underwent phase transitions characteristic of cholesterol esters, as observed by polarizing microscopy. The mean cholesteric to isotropic phase transition temperature of the anisotropic droplets was 41.3 degrees C. These data indicate that the anisotropic droplets of the oval fat bodies were composed of cholesterol esters acylated largely with monounsaturated fatty acids consistent with cellular origin. The noncellular fractions were subjected to ultracentrifugation (48,000 X g for 2 hours) and then separated into supernate and infranate fractions. The supernate fractions contained minor amounts of lipid, except in two cases of massive lipiduria. In these two cases, the supernate fractions contained many individual anisotropic droplets with lipid composition nearly identical to their cellular fractions. The dispersed lipid of the infranate fractions were invisible by microscope. Nine of the 11 infranate fractions revealed an alpha migrating lipid band by agarose gel electrophoresis. For urine specimens with anisotropic droplets, the infranate fractions contained cholesterol ester fatty acids that were less saturated (as measured by gas-liquid chromatography) than the cellular fractions. Thus, the cholesterol esters of the infranate fractions were derived from a different source (probably high-density lipoproteins) than the cholesterol esters of the cellular fractions.