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ToxSci Advance Access originally published online on January 27, 2006
Toxicological Sciences 2006 91(1):218-226; doi:10.1093/toxsci/kfj119
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© The Author 2006. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Oxidative Ability and Toxicity of n-Hexane Insoluble Fraction of Diesel Exhaust Particles

Hirotoshi Shima*, Eiko Koike*, Ritsuko Shinohara{dagger} and Takahiro Kobayashi{ddagger},1

* PM2.5 and DEP Research Project, National Institute for Environmental Studies, Onogawa 16-2, Tsukuba, Ibaraki 305-8506, Japan; {dagger} Department of Biomolecular Science, Faculty of Science, Toho University, Funabashi, Chiba 274-8510, Japan; and {ddagger} Environmental Health Sciences Division, National Institute for Environmental Studies, Tsukuba, Ibaraki 305-8506, Japan

Received November 14, 2005; accepted January 14, 2006

Diesel exhaust particles (DEP) are known to induce adverse biological responses such as inflammation of the airway. However, the relationship between the chemical characteristics of organic compounds adsorbed on DEP and their biological effects is not yet fully understood. In this study, the dichloromethane-soluble fraction (DMSF) from DEP was fractionated into its n-hexane-soluble fraction (n-HSF) and n-hexane-insoluble fraction (n-HISF). Using these DEP fractions, we designed the present studies to elucidate (1) chemical characteristics, (2) biological characteristics, and (3) the relationship between the chemical and the biological characteristics of these DEP fractions. Dithiothreitol (DTT) assay, Fourier transform-infrared (FT-IR) spectroscopy, proton nuclear magnetic resonance (1H-NMR) spectroscopy, and gas chromatography-mass spectrometry (GC-MS) were used to characterize their chemical properties. Heme oxygenase-1 (HO-1) protein expression, viability of rat alveolar type II epithelial cell line (SV40T2), and inflammatory cell infiltration into the peritoneal cavity of BALB/c mice were evaluated as markers of oxidative stress, cytotoxicity, and inflammatory response, respectively. The oxidative ability of the DEP fractions was n-HISF > DMSF > n-HSF. IR, 1H-NMR, and GC-MS spectra showed that n-HISF was mainly composed of compounds having many functional groups related to oxygenation, such as hydroxyl and carbonyl groups. The relative strength of HO-1 protein expression, cytotoxicity, and inflammatory responses was also n-HISF > DMSF > n-HSF. All of the n-HISF-induced biological activities were decreased by reduction with N-acetyl-L-cysteine (NAC). These results suggest that n-HISF has high oxidative ability and many functional groups related to oxygenation and that this ability strongly contributes to the induction of oxidative stress, cytotoxicity, and inflammatory response.

Key Words: diesel exhaust particles; n-hexane-insoluble fraction (n-HISF); oxidative ability; oxidative stress; cytotoxicity; inflammation.


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