Dobbs L G, Gonzalez R, Matthay M A, Carter E P, Allen L, Verkman A S
Department of Medicine, University of California, San Francisco, CA, 94143, USA.
Proc Natl Acad Sci U S A. 1998 Mar 17;95(6):2991-6. doi: 10.1073/pnas.95.6.2991.
Water permeability measured between the airspace and vasculature in intact sheep and mouse lungs is high. More than 95% of the internal surface area of the lung is lined by alveolar epithelial type I cells. The purpose of this study was to test whether osmotic water permeability (Pf) in type I alveolar epithelial cells is high enough to account for the high Pf of the intact lung. Pf measured between the airspace and vasculature in the perfused fluid-filled rat lung by the pleural surface fluorescence method was high (0.019 +/- 0.004 cm/s at 12 degrees C) and weakly temperature-dependent (activation energy 3.7 kcal/mol). To resolve the contributions of type I and type II alveolar epithelial cells to lung water permeability, Pf was measured by stopped-flow light scattering in suspensions of purified type I or type II cells obtained by immunoaffinity procedures. In response to a sudden change in external solution osmolality from 300 to 600 mOsm, the volume of type I cells decreased rapidly with a half-time (t1/2) of 60-80 ms at 10 degrees C, giving a plasma membrane Pf of 0.06-0.08 cm/s. Pf in type I cells was independent of osmotic gradient size and was weakly temperature-dependent (activation energy 3.4 kcal/mol). In contrast, t1/2 for type II cells in suspension was much slower, approximately 1 s; Pf for type II cells was 0.013 cm/s. Vesicles derived from type I cells also had a very high Pf of 0.06-0.08 cm/s at 10 degrees C that was inhibited 95% by HgCl2. The Pf in type I cells is the highest measured for any mammalian cell membrane and would account for the high water permeability of the lung.
在完整的绵羊和小鼠肺中,气腔与脉管系统之间测得的水通透性很高。肺内表面积的95%以上由I型肺泡上皮细胞覆盖。本研究的目的是测试I型肺泡上皮细胞中的渗透水通透性(Pf)是否足够高,以解释完整肺的高Pf。通过胸膜表面荧光法在灌注有液体的大鼠肺的气腔与脉管系统之间测得的Pf很高(12℃时为0.019±0.004 cm/s),且对温度的依赖性较弱(活化能为3.7 kcal/mol)。为了确定I型和II型肺泡上皮细胞对肺水通透性的贡献,通过免疫亲和程序获得的纯化I型或II型细胞悬液中的Pf,采用停流光散射法进行测量。当外部溶液渗透压从300突然变为600 mOsm时,在10℃下,I型细胞的体积迅速减小,半衰期(t1/2)为60 - 80毫秒,质膜Pf为0.06 - 0.08 cm/s。I型细胞中的Pf与渗透梯度大小无关,且对温度的依赖性较弱(活化能为3.4 kcal/mol)。相比之下,悬浮的II型细胞的t1/2要慢得多,约为1秒;II型细胞的Pf为0.013 cm/s。源自I型细胞的囊泡在10℃时也具有非常高的Pf,为0.06 - 0.08 cm/s,HgCl2可抑制其95%。I型细胞中的Pf是所测量的任何哺乳动物细胞膜中最高的,这可以解释肺的高水通透性。