The kinetics of the pulmonary endothelial uptake of serotonin (5-HT) were evaluated in isolated dog lung lobes using three methods. In method A serotonin was infused at various constant rates to provide a range of capillary concentrations that included Km. The arterial and venous concentrations measured by high-performance liquid chromatography were then used to determine the effect of concentration on the rate of 5-HT uptake. In method B trace doses of 5-[3H]HT and a reference indicator (indocyanine green dye) were injected during each constant infusion of unlabeled 5-HT to provide a measure of unidirectional 5-HT uptake at each background concentration. In method C boluses containing different amounts of unlabeled 5-HT, along with the 5-[3H]HT and the dye, were injected such that each bolus resulted in a range of concentrations and provided a measure of the unidirectional uptake at each concentration. Each method provided the data needed to calculate the maximum uptake rate (Vmax) and the concentration at Vmax/2 (Km), assuming that the uptake kinetics can be represented by the Michaelis-Menten equation. However, the mathematical model underlying each method involved different assumptions about the returning flux of the 5-HT which entered the endothelial cell and the heterogeneity of vascular transit times. The results obtained, considered in light of the different assumptions involved, indicate that all three methods can provide reasonable estimates of the mass transfer kinetic constants if the constant infusions of 5-HT are of short duration and/or the boluses are adequately dispersed prior to reaching the capillary bed.

1 Pulmonary prostacyclin (PGI2) biosynthesis was evaluated in relation to endothelial integrity before and after complement activation in isolated plasma-perfused lung lobes of the dog. 2 The plasma was activated with zymosan (ZAP, n = 4), yeast cells (YAP, n = 4) or yeast with 3 yM indomethacin (Indo + YAP, n = 3). Immunoreactive 6-oxo-prostaglandin F a (i-6-oxo-PGF J and thromboxane B2 (iTXB2) were measured to monitor PGI2 and TXA2 biosynthesis.3 The kinetic parameters Km and V,,. of 5-hydroxytryptamine (5-HT) uptake were calculated on the basis of multiple indicator diffusion data to evaluate endothelial integrity. 4 YAP and ZAP induced a biphasic increase of the arterial perfusion pressure. The immediate pressure peak was partly mediated by TXA2 and the TXB2 was subsequently cleared by the lung.5 The apparent V.,x of 5-HT uptake remained constant throughout the experiment. Thus, complement activation did not affect the number of endothelial 5-HT carrier sites available to the perfusate. 6 The apparent Km of 5-HT uptake was enhanced in 9 lungs exposed to activated plasma complement for 20 min. This decreased affinity for 5-HT probably reflects endothelial injury. It was transient as the apparent Km had returned to the baseline value after 60 min. 7 PGI2 clearance and biosynthesis were virtually absent in the control period. PGI2 formation increased drastically after infusion of ZAP or YAP and was proportional to the endothelial injury expressed as elevated Km or pulmonary oedema. Thus, PGI2 biosynthesis might be a marker of severe endothelial distress.

Prostacyclin (PGI2) formation and the saturable uptake of 5-hydroxytryptamine (5HT) were studied as indices of endothelial integrity in isolated lungs. 5HT uptake was characterized by its kinetic parameters Km and Vmax. These were calculated from multiple indicator dilution data on the basis of an organ model of 5HT uptake. Perfused dog lung lobes were exposed to plasma activated with yeast (YAP) or zymosan (ZAP). YAP induced a transient elevation of Km. This increase probably reflects endothelial injury. Vmax remained unchanged, suggesting that the perfused endothelial surface remained stable. PGI2 biosynthesis was negligible in the control period, but started immediately after exposure to ZAP or YAP. It was proportional to the transient elevation of Km and to the pulmonary oedema. The data suggest that PGI2 might be a marker of severe endothelial distress.