Zeidel M L, Kikeri D, Silva P, Burrowes M, Brenner B M
Renal Division, Brigham and Women's Hospital, Boston, Massachusetts 02115.
J Clin Invest. 1988 Sep;82(3):1067-74. doi: 10.1172/JCI113663.
The inner medullary collecting duct (IMCD) effects net sodium reabsorption under the control of volume regulatory hormones, including atrial natriuretic peptides (ANP). These studies examined the mechanisms of sodium transport and its regulation by ANP in fresh suspensions of IMCD cells. Sodium uptake was inhibited by amiloride but insensitive to furosemide, bu-metanide, and hydrochlorthiazide. These results are consistent with uptake mediated by a sodium channel or Na+/H+ exchange. To determine the role of sodium channels, cells were hyperpolarized by preincubation in high potassium medium followed by dilution into potassium-free medium. Membrane potential measurements using the cyanine dye, Di(S)-C3-5 verified a striking hyperpolarization of IMCD cells using this protocol. Hyperpolarization increased the apparent initial rate of sodium uptake fourfold. Amiloride and ANP inhibited potential-stimulated sodium uptake 73% and 65%, respectively; the two agents together were not additive. Addition of 5 mM sodium to hyperpolarized cells resulted in a significant amiloride-sensitive depolarization. Half-maximal inhibition of potential-driven sodium uptake occurred at 3 X 10(-7) M amiloride, and 5 X 10(-11) M ANP. We conclude that sodium enters IMCD cells via a conductive, amiloride-sensitive sodium channel, which is regulated by ANP. ANP inhibition of luminal sodium entry in the IMCD appears to contribute to the marked natriuretic effect of this hormone in vivo.
内髓集合管(IMCD)在包括心房利钠肽(ANP)在内的容量调节激素的控制下实现钠的净重吸收。这些研究检测了IMCD细胞新鲜悬液中钠转运及其受ANP调节的机制。阿米洛利可抑制钠摄取,但对呋塞米、布美他尼和氢氯噻嗪不敏感。这些结果与由钠通道或Na+/H+交换介导的摄取一致。为了确定钠通道的作用,先将细胞置于高钾培养基中预孵育使其超极化,然后再稀释到无钾培养基中。使用花菁染料Di(S)-C3-5进行的膜电位测量证实,采用该方案时IMCD细胞出现显著超极化。超极化使钠摄取的表观初始速率提高了四倍。阿米洛利和ANP分别抑制电位刺激的钠摄取73%和65%;二者共同作用时无叠加效应。向超极化细胞中加入5 mM钠会导致阿米洛利敏感的显著去极化。电位驱动的钠摄取被半数抑制时,阿米洛利的浓度为3×10(-7) M,ANP的浓度为5×10(-11) M。我们得出结论,钠通过一种对阿米洛利敏感的传导性钠通道进入IMCD细胞,该通道受ANP调节。ANP对IMCD管腔钠进入的抑制作用似乎是该激素在体内显著利钠作用的原因。