cDNA microarray analysis of rat alveolar epithelial cells following exposure to organic extract of diesel exhaust particles.
Diesel exhaust particles (DEP) induce pulmonary diseases including asthma and chronic bronchitis. Comprehensive evaluation is required to know the mechanisms underlying the effects of air pollutants including DEP on lung diseases. Using a cDNA microarray, we examined changes in gene expression in SV40T2 cells, a rat alveolar type II epithelial cell line, following exposure to an organic extract of DEP. We identified candidate sensitive genes that were up- or down-regulated in response to DEP. The cDNA microarray analysis revealed that a 6-h exposure to the DEP extract (30 microg/ml) increased (>2-fold) the expression of 51 genes associated with drug metabolism, antioxidation, cell cycle/proliferation/apoptosis, coagulation/fibrinolysis, and expressed sequence tags (ESTs), and decreased (<0.5-fold) that of 20 genes. In the present study, heme oxygenase (HO)-1, an antioxidative enzyme, showed the maximum increase in gene expression; and type II transglutaminase (TGM-2), a regulator of coagulation, showed the most prominent decrease among the genes. We confirmed the change in the HO-1 protein level by Western blot analysis and that in the enzyme activity of TGM-2. The organic extract of DEP increased the expression of HO-1 protein and decreased the enzyme activity of TGM-2. Furthermore, these effects of DEP on either HO-1 or TGM-2 were reduced by N-acetyl-l-cysteine (NAC), thus suggesting that oxidative stress caused by this organic fraction of DEP may have induced these cellular responses. Therefore, an increase in HO-1 and a decrease in TGM-2 might be good markers of the biological response to organic compounds of airborne particulate substances.
Koike
Eiko
E
Particulate Matter (PM2.5) and Diesel Exhaust Particles (DEP) Research Project, National Institute for Environmental Studies, Tsukuba, Ibaraki, 305-8506, Japan. ekoike@nies.go.jp
Hirano
Seishiro
S
Furuyama
Akiko
A
Kobayashi
Takahiro
T
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics*; D015536:Down-Regulation; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism; D006419:Heme Oxygenase (Decyclizing) / Q000378:metabolism; D051547:Heme Oxygenase-1; D020411:Oligonucleotide Array Sequence Analysis; D018384:Oxidative Stress / Q000187:drug effects; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011650:Pulmonary Alveoli / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D011503:Transglutaminases / Q000378:metabolism; D015854:Up-Regulation; D001335:Vehicle Emissions / Q000633:toxicity*
D018076:DNA, Complementary; C000717609:Tgm2 protein, rat; D001335:Vehicle Emissions; D006419:Heme Oxygenase (Decyclizing); D051547:Heme Oxygenase-1; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases