| Autor: |
Spokas, Eric G.1 espokas@rider.edu, Harshman, Scott2, Cohen, Glenn M.3, Chen Jiang3, Levine, Jaime M.4, Rodriguez, Ana R.4, Foglein, Jon2, Spur, Bernd W.4 |
| Předmět: |
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| Zdroj: |
Environmental Toxicology & Chemistry. Jul2008, Vol. 27 Issue 7, p1569-1575. 7p. 3 Color Photographs, 1 Black and White Photograph, 1 Chart, 2 Graphs. |
| Abstrakt: |
The aim of the present study was to evaluate oxidative injury in gill pavement cells (GPCs) from fathead minnow (Pimephales promelas) using F2-isoprostane (F2-iP) release as an index of lipid peroxidation. Cells were isolated from pooled gill tissue by collagenase treatment, mechanical sieving, and Percoll® density gradient centrifugation. Baseline levels of 8-epi-prostaglandin F2α (8-epi-PGF2α) were measured by incubating GPCs in physiological buffer (106 cells/ml) and enzyme immunoassay. After 60 min, the amount of immunoreactive 8-epi-PGF2α (ir8-epi-PGF2α) in control medium ranged from 1,374 to 5,515 pg/ml. Lead nitrate, 0.6 to 120 μM, did not influence ir8-epi-PGF2α release, whereas FeCl3 stimulated release at 500 μM but not at 5 μM. Incubation medium was extracted for acidic lipids and analyzed by liquid chromatography/mass spectrometry/electrosprayionization. A compound in the medium exhibited a retention time on reverse-phase high-performance liquid chromatography nearly identical to that of synthetic 8-epi-PGF2α. The mass spectrum taken from the total ion chromatogram from 14.8 to 15.1 min contained a prominent ion at m/z 353, as expected for the molecular ion of 8-epi-PGF2α. Similar results were obtained with tissue subjected to base hydrolysis. Mass spectra of extracted ion chromatograms obtained with gill extracts and authentic standard showed a close correspondence of fragment ions, providing definitive evidence for production and storage of F2-iPs by fish gills. In summary, F2-iP release occurs during lipid peroxidation injury to fish gill epithelium, and its measurement may facilitate aquatic toxicology studies of metallic and nonmetallic contaminants. [ABSTRACT FROM AUTHOR] |
| Databáze: |
GreenFILE |
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