Shafrir E, Levy E, Deckelbaum R
Department of Biochemistry, Hadassah University Hospital, Jerusalem, Israel.
Nephron. 1990;56(1):24-9. doi: 10.1159/000186095.
Lipoprotein excretion was investigated in the urine of hyperlipidemic rats with nephrotic syndrome induced by aminonucleoside of puromycin. Incubation with phosphatidylcholine liposomes was employed to float apoproteins not bound to lipids, by ultracentrifugation at d = 1.21 g/ml. On ultracentrifugation of whole, untreated urine, the amount of protein floated was 6-fold greater in nephrotic vs. control rats and consisted mainly of HDL-like particles. Sodium dodecylsulfate-polyacrylamide gel electrophoresis showed that control urine contained apoproteins A-I, A-II, E and traces of C, whereas in the nephrotic urine apo-E and a large amount of apo-C was found. Addition of liposomes to the ultracentrifugal d = 1.21 g/ml infranate and reflotation at the same density resulted only in slight increment in the floated apoproteins, mainly C and A-I. Addition of liposomes to the whole urine and centrifugation at d = 1.21 g/ml also did not produce a greater yield in the floated apoproteins of control or nephrotic urine. These results indicated that the urine is virtually devoid of lipid-free apoproteins and those floated from both the nephrotic and control urine are complexed with lipids. The plasma VLDL + LDL fraction of nephrotic rats, though increased in quantity, did not differ markedly in composition from that of control rats. The HDL, approximately 3-fold elevated in nephrotic rats, were poorer in esterified cholesterol and richer in phospholipids. Relative to plasma HDL, the nephrotic urine HDL were protein-rich and phospholipid-poor and appeared to be larger in particle size as suggested by the lower estimated specific volume. The modified plasma HDL in nephrosis may have a pathophysiological implication.
在嘌呤霉素氨基核苷诱导的高脂血症肾病综合征大鼠的尿液中研究了脂蛋白排泄情况。通过在密度d = 1.21 g/ml下超速离心,用磷脂酰胆碱脂质体孵育以漂浮未与脂质结合的载脂蛋白。对未经处理的全尿进行超速离心时,肾病大鼠尿液中漂浮的蛋白量比对照大鼠高6倍,且主要由类高密度脂蛋白颗粒组成。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示,对照尿液中含有载脂蛋白A-I、A-II、E和微量的C,而肾病尿液中发现了载脂蛋白E和大量的载脂蛋白C。向密度d = 1.21 g/ml的超速离心下层液中添加脂质体并在相同密度下再次漂浮,仅使漂浮的载脂蛋白略有增加,主要是C和A-I。向全尿中添加脂质体并在密度d = 1.21 g/ml下离心,也未使对照或肾病尿液中漂浮的载脂蛋白产量增加。这些结果表明,尿液中实际上不含无脂质的载脂蛋白,从肾病和对照尿液中漂浮的载脂蛋白都与脂质结合。肾病大鼠的血浆极低密度脂蛋白+低密度脂蛋白部分,尽管数量增加,但其组成与对照大鼠相比无明显差异。肾病大鼠中高密度脂蛋白升高约3倍,其酯化胆固醇含量较低,磷脂含量较高。相对于血浆高密度脂蛋白,肾病尿液中的高密度脂蛋白富含蛋白质且磷脂含量低,并且根据较低的估计比容表明其颗粒尺寸似乎更大。肾病中修饰的血浆高密度脂蛋白可能具有病理生理学意义。