Preparation of human lung tissue from cigarette smokers for analysis by electron spin resonance spectroscopy


This chapter describes an electron spin resonance (ESR) study of freeze-dried tissue from smokers. Frozen whole lung specimens are obtained from a light and from a heavy cigarette smoker. The lungs from the light smoker show only a light mottling, while tissues from the smoker show extensive areas of dark rust-brown pigmentation surrounded by tissue that appears similar in color to that of normal lung tissue. The ESR analyses are carried out using a Brucker ESR spectrometer model ER/100. The results show that the lungs, and especially the alveolar macrophages, of cigarette smokers contain high concentrations of iron. The ESR results suggest that this iron either is in ferritin or hemosiderin or is very similar to the form of iron occurring in these storage proteins. The accumulation of iron, especially if it is ferritin- or hemosiderin-like, in the lungs of smokers may have an important impact on the level of oxidative stress to which the lungs of smokers are exposed. The iron in ferritin is bound in the +3 oxidation state and is unreactive; however, this iron can be easily released from the ferritin core as ferrous iron by the action of reducing agents and, thereby, be made available to initiate oxidative damage such as lipid peroxidation. The data presented in the chapter leads to following hypothesis that cigarette smoke produces high concentrations of oxidizing radicals in a microdomain in the lung, leading to microhemorrhaging. © 1990, Elsevier Inc. All rights reserved.

Publication Title

Methods in Enzymology