Ys. Bai et al., The cytotoxic effects of diesel exhaust particles on human pulmonary artery endothelial cells in vitro: Role of active oxygen species, FREE RAD B, 30(5), 2001, pp. 555-562
Diesel exhaust particles (DEP) have been proved to induce serious pulmonary
injury, among which lethal pulmonary edema has been assumed to be mediated
by vascular endothelial cell damage. In the present study, we investigated
the cytotoxic mechanism of DEP on human pulmonary artery endothelial cells
focusing on the role of active oxygen species. Endothelial cell viability
was assessed by WST-8, a novel tetrazolium salt. Nitric oxide (NO) producti
on was measured by using a new fluorescence indicator, diaminofluorescein-2
(DAF-2). Organic compounds in DEP were extracted by dichloromethane and me
thanol. DEP-extracts damaged endothelial cells under both subconfluent and
confluent conditions. The DEP-extract-induced cytotoxicity was markedly red
uced by treatment with SOD, catalase, N-(2-mercaptopropionyl)-glycine (MPG)
, or ebselen (a selenium-containing compound with glutathione peroxidase-li
ke activity). Thus superoxide, hydrogen peroxide, and other oxygen-derived
free radicals are likely to be implicated in DEP-extract-induced endothelia
l cell damage. Moreover, L-NAME and L-NMA, inhibitors of NO synthase, also
attenuated DEP-extract-induced cytotoxicity, while sepiapterin, the precurs
or of tetrahydrobiopterin (BH4, a NO synthase cofactor) interestingly enhan
ced DEP-extract-induced cell damage. These findings suggest that NO is also
involved in DEP-extract-mediated cytotoxicity, which was confirmed by dire
ct measurement of NO production. These active oxygen species, including per
oxynitrite, may explain the mechanism of endothelial cell damage upon DEP e
xposure during the early stage. (C) 2001 Elsevier Science Inc.