The processing of exogenous cholesterol in the alveolar compartment of the rat lung

Abstract

We have examined the esterification of [3H]cholesterol following the intratracheal instillation of a tracer amount into the isolated rat lung perfused with Krebs bicarbonate containing 4.5% albumin. At 5, 30 and 60 min after instillation, lungs were lavaged at 2 degrees C with 3 x 10 ml of 0.15 M NaCl, each volume instilled and withdrawn three times. Each lung was lavaged at only one time point. The saline recovered was centrifuged at 150 g (5 min) to sediment the macrophage-rich fraction, leaving the surfactant in the supernatant. The amounts and specific activity of cholesterol and cholesteryl ester were measured following isolation by high performance liquid chromatography of the free cholesterol and the hydrolyzed ester-derived cholesterol. There was a rapid fall in [3H]cholesterol in the surfactant fraction, accompanied by a reciprocal increase in [3H]cholesteryl ester in the macrophage-rich fraction, while the level of free [3H]cholesterol in that fraction remained very low. These data are consistent with exogenous cholesterol being rapidly esterified in the alveolus, and the ester then being cleared by the macrophages. We were unable to locate the actual site of esterification.