SMOKING

Long term smoking with age builds up excessive oxidative stress in bronchoalveolar lavage fluid

K Nagai¹, T Betsuyaku¹, T Kondo², Y Nasuhara¹, M Nishimura¹

¹ First Department of Medicine, Hokkaido University School of Medicine, Sapporo, Japan
² Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan

Correspondence to:
Dr T Betsuyaku
First Department of Medicine, Hokkaido University School of Medicine, N-15, W-7, Kita-ku, Sapporo, Japan 060-8638; bytomoko{at}med.hokudai.ac.jp

Background: Epithelial lining fluid plays a critical role in protecting the lung from oxidative stress, in which the oxidised status may change by ageing, smoking history, and pulmonary emphysema.

Methods: Bronchoalveolar lavage (BAL) was performed on 109 young and older subjects with various smoking histories. The protein carbonyls, total and oxidised glutathione were examined in BAL fluid.

Results: By Western blot analysis, the major carbonylated protein in the BAL fluid was sized at 68 kDa, corresponding to albumin. The amount of carbonylated albumin per mg total albumin in BAL fluid was four times higher in older current smokers and three times higher in older former smokers than in age matched non-smokers (p<0.0001, p = 0.0003, respectively), but not in young smokers. Total glutathione in BAL fluid was significantly increased both in young (p = 0.006) and older current smokers (p = 0.0003) compared with age matched non-smokers. In contrast, the ratio of oxidised to total glutathione was significantly raised (72%) only in older current smokers compared with the other groups. There was no significant difference in these parameters between older smokers with and without mild emphysema.

Conclusions: Oxidised glutathione associated with excessive protein carbonylation accumulates in the lung of older smokers with long term smoking histories even in the absence of lung diseases, but they are not significantly enhanced in smokers with mild emphysema.

Abbreviations: BAL, bronchoalveolar lavage; DNP, dinitrophenylhydrazine; ELF, epithelial lining fluid; FEV₁, forced expiratory volume in 1 second; GSH, reduced glutathione; GSSG, glutathione disulfide

Keywords: bronchoalveolar lavage; protein carbonyls; antioxidant; glutathione; smoking; ageing

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