Induction of apoptosis by glyoxal in human embryonic lung epithelial cell line L132.
Oxidative stress has been suggested to play a central role in the pathogenesis of lung fibrosis and lung epithelial cell apoptosis is considered to be a key event during fibrogenesis. Studies from various laboratories have indicated that metabolic conditions may initiate oxidative stress, thereby contributing to epithelial cell death. This study was designed to test the hypothesis that glyoxal, an intermediate product in the glycation reaction leading to advanced glycation end products (AGEs), may induce lung epithelial cell apoptosis. We investigated the in vitro effects of glyoxal on fetal human lung epithelial L132 cells. Immunocytochemical analysis of paraffin-embedded cells and fluorescence-activated cell sorter analysis revealed a dose-dependent accumulation of the glycoxidation product (epsilon)N-carboxymethyllysine (CML) in all compartments of the cell. It has been shown that CML modification of proteins may serve as an indicator for oxidative stress. To examine the role of apoptosis in epithelial lung cells we investigated glyoxal-dependent changes in pro- and antiapoptotic mediators bax and activated caspase-3, and galectin-3 and bcl-2, respectively. Increasing concentrations of glyoxal (50 to 400 microM) induced an increase in the number of apoptotic cells. The apoptotic changes were confirmed by transmission electron microscopy. Immunocytochemical analysis of treated cells revealed the presence of other AGEs such as pentosidine as well as products of lipid peroxidation.
Kasper
M
M
Institute of Anatomy and Institute of Pathology, Technical University of Dresden, Dresden, Germany. mkas@rcs.urz.tu-dresden.de
Roehlecke
C
C
Witt
M
M
Fehrenbach
H
H
Hofer
A
A
Miyata
T
T
Weigert
C
C
Funk
R H
RH
Schleicher
E D
ED
D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002460:Cell Line; D004789:Enzyme Activation; D004847:Epithelial Cells / Q000187:drug effects / Q000201:enzymology; D005434:Flow Cytometry; D006037:Glyoxal / Q000494:pharmacology*; D006801:Humans; D006868:Hydrolysis; D007150:Immunohistochemistry; D008168:Lung / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D008854:Microscopy, Electron
D006037:Glyoxal; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases