Silverman M
J Clin Invest. 1973 Oct;52(10):2486-94. doi: 10.1172/JCI107439.
The localization of disaccharidases in kidney has been studied by means of the multiple indicator dilution technique. A pulse injection of a solution containing Evans blue dye (plasma marker), creatinine (extracellular marker), and a (14)C-labeled disaccharide (lactose, sucrose, maltose, and alphaalpha-trehalose), is made into the renal artery of an anesthetized dog, and the outflow curves are monitored simultaneously from renal venous and urine effluents. Lactose and sucrose have an extracellular distribution. Trehalose and maltose remain extracellular from the postglomerular circulation. About 75% of filtered tracer maltose or trehalose is extracted by the luminal surface of the nephron. Thin-layer chromatography of urine samples shows that all of the excreted (14)C radiolabel is in the form of the injected disaccharide. Following the administration of phlorizin, all of the filtered radioactivity is recoverable in the urine, but chromatography of the urine samples now reveals that there is a significant excretion of [(14)C]glucose, approximating the amount previously extracted under control conditions (in the absence of phlorizin). It has been verified that no hydrolysis of maltose or trehalose to their constituent glucose subunits occurred during the transit of tracer between the point of injection (renal artery), and the point of filtration (glomerular basement membrane). Similarly, after addition of [(14)C]disaccharides to fresh urine there is no chromatographically recoverable [(14)C]glucose. It is concluded that there exist alpha-glucosidases with maltase and trehalase activity along the brush border of the proximal tubule and that these disaccharidases are located spatially superficial to the glucose transport receptors.
已通过多指示剂稀释技术研究了双糖酶在肾脏中的定位。将含有伊文思蓝染料(血浆标志物)、肌酐(细胞外标志物)和一种¹⁴C标记双糖(乳糖、蔗糖、麦芽糖和α,α-海藻糖)的溶液脉冲注入麻醉犬的肾动脉,并同时监测肾静脉和尿液流出液的流出曲线。乳糖和蔗糖具有细胞外分布。海藻糖和麦芽糖在肾小球后循环中仍处于细胞外。约75%滤过的示踪剂麦芽糖或海藻糖被肾单位的管腔表面摄取。尿样的薄层色谱分析表明,所有排泄的¹⁴C放射性标记均为注入双糖的形式。给予根皮苷后,所有滤过的放射性均可在尿液中回收,但尿样的色谱分析现在显示有大量的¹⁴C葡萄糖排泄,其数量接近先前在对照条件下(无根皮苷)摄取的量。已经证实,在示踪剂从注射点(肾动脉)到滤过点(肾小球基底膜)的转运过程中,麦芽糖或海藻糖没有水解为其组成的葡萄糖亚基。同样,在向新鲜尿液中添加¹⁴C双糖后,没有色谱可回收的¹⁴C葡萄糖。结论是,近端小管刷状缘存在具有麦芽糖酶和海藻糖酶活性的α-葡萄糖苷酶,并且这些双糖酶在空间上位于葡萄糖转运受体的表面。