Cheung P T, Hammerman M R
Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
Am J Physiol. 1988 May;254(5 Pt 2):F711-8. doi: 10.1152/ajprenal.1988.254.5.F711.
To define the mechanism by which glucose is transported across the basolateral membrane of the renal proximal tubular cell, we measured D-[14C]glucose uptake in basolateral membrane vesicles from rabbit kidney. Na+-dependent D-glucose transport, demonstrable in brush-border vesicles, could not be demonstrated in basolateral membrane vesicles. In the absence of Na+, the uptake of D-[14C]glucose in basolateral vesicles was more rapid than that of L-[3H]glucose over a concentration range of 1-50 mM. Subtraction of the latter from the former uptakes revealed a saturable process with apparent Km of 9.9 mM and Vmax of 0.80 nmol.mg protein-1.s-1. To characterize the transport component of D-glucose uptake in basolateral vesicles, we measured trans stimulation of 2 mM D-[14C]glucose entry in the absence of Na+. Trans stimulation could be effected by preloading basolateral vesicles with D-glucose, 2-deoxy-D-glucose, or 3-O-methyl-D-glucose, but not with L-glucose or alpha-methyl-D-glucoside. Trans-stimulated D-[14C]glucose uptake was inhibited by 0.1 mM phloretin or cytochalasin B but not phlorizin. In contrast, Na+-dependent D-[14C]glucose transport in brush-border vesicles was inhibited by phlorizin but not phloretin or cytochalasin B. Our findings are consistent with the presence of a Na+-independent D-glucose transporter in the proximal tubular basolateral membrane with characteristics similar to those of transporters present in nonepithelial cells.
为了确定葡萄糖跨肾近端小管细胞基底外侧膜转运的机制,我们测量了兔肾基底外侧膜囊泡中D-[14C]葡萄糖的摄取。在刷状缘囊泡中可证明的Na+依赖性D-葡萄糖转运,在基底外侧膜囊泡中无法证明。在没有Na+的情况下,在1-50 mM的浓度范围内,基底外侧囊泡中D-[14C]葡萄糖的摄取比L-[3H]葡萄糖的摄取更快。从前者摄取中减去后者摄取揭示了一个可饱和过程,其表观Km为9.9 mM,Vmax为0.80 nmol·mg蛋白-1·s-1。为了表征基底外侧囊泡中D-葡萄糖摄取的转运成分,我们在没有Na+的情况下测量了2 mM D-[14C]葡萄糖进入的反式刺激。反式刺激可以通过用D-葡萄糖、2-脱氧-D-葡萄糖或3-O-甲基-D-葡萄糖预加载基底外侧囊泡来实现,但不能用L-葡萄糖或α-甲基-D-葡萄糖苷。反式刺激的D-[14C]葡萄糖摄取被0.1 mM根皮素或细胞松弛素B抑制,但不被根皮苷抑制。相反,刷状缘囊泡中Na+依赖性D-[14C]葡萄糖转运被根皮苷抑制,但不被根皮素或细胞松弛素B抑制。我们的发现与近端小管基底外侧膜中存在一种Na+非依赖性D-葡萄糖转运体一致,其特征与非上皮细胞中存在的转运体相似。