Studies on the mechanisms of active ion fluxes across alveolar epithelial cell monolayers

Abstract

To investigate the cell physiologic and biological properties of the alveolar epithelium, we studied rat alveolar epithelial cell monolayers grown on permeable supports in primary culture. Type II alveolar epithelial cells were disaggregated using elastase, and partially purified on a discontinuous metrizamide gradient. These isolated cells were plated onto tissue culture-treated Nuclepore membrane filters at 1.5×106 cells/cm2 and maintained in a humidified incubator (5% CO2 in air, 37° C). After 2 days in culture, the bathing media on both sides of the cell monolayers were changed to fresh culture medium, thus removing nonadherent cells (mostly leukocytes). These monolayers exhibit a high transmonolayer resistance (>2000 Ω-cm2) and actively transport ions. Radionuclide flux studies indicate that Na+ is the predominant ionic species absorbed actively under baseline conditions, accounting for about 80% of the total active ion transport. Cl− seems to be passively transported across the epithelium. However, when the epithelium is exposed to a beta-agonist (terbutaline), active absorption of Na+ is increased and active absorption of Cl− occurs. Although it is clear that both active Na+ and Cl− transport are dependent on Na+/K+-ATPase activity, and that Na+ enters cells predominantly through channels, the specific mechanisms by which Cl− enters and exits the alveolar epithelial cells remain unclear. The stimulated reabsorption of Na+ and Cl− may be important in helping to remove excess fluid from alveolar air spaces in the lung.