Tosco M, Orsenigo M N, Esposito G, Faelli A
Dipartimento di Fisiologia e Biochimica Generali, Università di Milano, Italy.
Cell Biochem Funct. 1988 Jul;6(3):155-64. doi: 10.1002/cbf.290060303.
The basolateral membrane of the jejunal enterocyte of the rat was separated by self-orienting Percoll-gradient centrifugation and further purified from brush border contamination. Pellets were analysed for Mg-, Na- and (Na,K)-ATPase activities. The uptake of 0.02 M NaCl was also followed by the rapid micro-filtration technique. Transintestinal transport of fluid and electrolytes, and cell water, Na and K were determined in the in vitro everted and incubated jejunum. There is ouabain-insensitive Na-ATPase in addition to the well-known (Na,K)-ATPase in the basolateral membrane. These are differently inhibited by furosemide and ethacrynate. Na uptake by osmotically active basolateral membrane vesicles is enhanced by ATP and a further enhancement is obtained if there is intravesicular K. The ATP effect is inhibited differently by strophanthidin, furosemide and ethacrynate. In the everted sac preparation, transintestinal transport of Na and fluid still occurs when the Na/K pump is totally inhibited by ouabain. These experimental results suggest that there is also a ouabain-insensitive Na pump, different from the Na/K pump, in the basolateral membrane.
通过自定向Percoll梯度离心法分离大鼠空肠肠上皮细胞的基底外侧膜,并进一步从刷状缘污染中纯化。对沉淀进行镁、钠和(钠,钾)-ATP酶活性分析。还采用快速微滤技术跟踪0.02M氯化钠的摄取。在体外翻转并孵育的空肠中测定液体、电解质以及细胞内水、钠和钾的跨肠运输。除了基底外侧膜中众所周知的(钠,钾)-ATP酶外,还有哇巴因不敏感的钠-ATP酶。速尿和利尿酸对它们的抑制作用不同。有活性的基底外侧膜囊泡对钠的摄取可被ATP增强,如果囊泡内有钾则进一步增强。毒毛花苷、速尿和利尿酸对ATP的作用有不同的抑制作用。在翻转囊泡制备中,当钠/钾泵被哇巴因完全抑制时,钠和液体的跨肠运输仍会发生。这些实验结果表明,基底外侧膜中还存在一种不同于钠/钾泵的哇巴因不敏感钠泵。