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在已建立的肾细胞系OK中鉴定近端肾小管转运功能。

Identification of proximal tubular transport functions in the established kidney cell line, OK.

作者信息

Malström K, Stange G, Murer H

出版信息

Biochim Biophys Acta. 1987 Aug 20;902(2):269-77. doi: 10.1016/0005-2736(87)90305-1.

Abstract

OK cells, derived from an American opossum kidney, were analyzed for proximal tubular transport functions. In monolayers, L-glutamate, L-proline, L-alanine, and alpha-methyl-glucopyranoside (alpha-methyl D-glucoside) were accumulated through Na+-dependent and Na+-independent transport pathways. D-Glucose and inorganic sulfate were accumulated equally well in the presence or absence of Na+. Influx of inorganic phosphate was only observed in the presence of Na+. Na+/alpha-methyl D-glucoside uptake was preferentially inhibited by phlorizin and D-glucose uptake by cytochalasin B. An amiloride-sensitive Na+-transport was also identified. In isolated apical vesicles (enriched 8-fold in gamma-glutamyltransferase), L-glutamate, L-proline, L-alanine, alpha-methyl D-glucoside and inorganic phosphate transport were stimulated by an inwardly directed Na+-gradient as compared to an inwardly directed K+-gradient. L-Glutamate transport required additionally intravesicular K+. D-Glucose transport was similar in the presence of a Na+- and a K+-gradient. Na+/alpha-methyl D-glucoside uptake was inhibited by phlorizin whereas cytochalasin B had no effect on Na+/D-glucose transport. An amiloride-sensitive Na+/H+ exchange mechanism was also found in the apical vesicle preparation. It is concluded that the apical membrane of OK cells contains Na+-coupled transport systems for amino acids, hexoses, protons and inorganic phosphate. D-Glucose appears a poor substrate for the Na+/hexose transport system.

摘要

对源自美洲负鼠肾脏的OK细胞进行了近端肾小管转运功能分析。在单层细胞中,L-谷氨酸、L-脯氨酸、L-丙氨酸和α-甲基-D-吡喃葡萄糖苷(α-甲基D-葡萄糖苷)通过Na⁺依赖性和Na⁺非依赖性转运途径进行积累。无论有无Na⁺,D-葡萄糖和无机硫酸盐的积累情况都一样好。无机磷酸盐的内流仅在有Na⁺存在时才观察到。Na⁺/α-甲基D-葡萄糖苷的摄取优先受到根皮苷的抑制,而D-葡萄糖的摄取则受到细胞松弛素B的抑制。还鉴定出一种氨氯地平敏感的Na⁺转运。在分离的顶端小泡(γ-谷氨酰转移酶富集8倍)中,与内向K⁺梯度相比,内向Na⁺梯度刺激了L-谷氨酸、L-脯氨酸、L-丙氨酸、α-甲基D-葡萄糖苷和无机磷酸盐的转运。L-谷氨酸的转运还需要囊泡内的K⁺。在存在Na⁺梯度和K⁺梯度时,D-葡萄糖的转运情况相似。根皮苷抑制Na⁺/α-甲基D-葡萄糖苷的摄取,而细胞松弛素B对Na⁺/D-葡萄糖的转运没有影响。在顶端小泡制剂中还发现了一种氨氯地平敏感的Na⁺/H⁺交换机制。得出的结论是,OK细胞的顶端膜含有用于氨基酸、己糖、质子和无机磷酸盐的Na⁺偶联转运系统。D-葡萄糖似乎不是Na⁺/己糖转运系统的良好底物。

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