Kinetics and cellular localisation of putrescine uptake in human lung tissue

Abstract

Background: The polyamines (putrescine, spermidine, and spermine) are involved in cellular growth, proliferation, and differentiation. In the lungs of various species, polyamines are accumulated by an active uptake system which also mediates the uptake of cystamine and paraquat. In the rat lung putrescine uptake has been shown to be cell-specific, occurring predominantly in the alveolar epithelium. The aim of this study was to characterise the uptake of putrescine in human lung.

Methods: Lung tissue was obtained from 31 patients undergoing surgery for lung cancer. Slices (0.7 mm thick) from non-tumour containing lung parenchyma were incubated for 15-60 minutes in Krebs-Ringer phosphate buffer with various concentrations of putrescine (2.5 to 80 mumol/l) containing 0.1 microCi [1,4-14C]-putrescine. Uptake was assessed from tissue radioactivity. For autoradiographic imaging, slices were incubated for 30 minutes with 2.5 mumol/l putrescine containing 2.5 mCi [1,4n-3H]-putrescine.

Results: The accumulation of [14C]-putrescine into slices was time-dependent and energy-dependent, and obeyed saturation kinetics, with mean calculated values for Vmax (maximal rate of uptake) of 414 nmol/g/hour and for Km (medium concentration at which the rate of uptake is half Vmax) of 7.2 mumol/l, with a large interindividual variation. Competitive inhibition was observed on incubation with cystamine, which appears to have a high affinity for the uptake system since its calculated Ki (concentration of inhibitor at which the Km is doubled) was 3.2 mumol/l. Ultrastructural autoradiography showed labelling over both type I and type II cells of the alveolar epithelium, but not over the endothelium or any cells of the interstitium. Alveolar macrophages were also devoid of label.

Conclusions: These results show that the human lung possesses an active uptake system for putrescine, and probably also cystamine, which is located in both cell types of the alveolar epithelium. These findings may be used to develop tests for the assessment of the alveolar epithelium.