EROD activities in a primary cell culture of grass carp (Ctenopharyngodon idellus) hepatocytes exposed to polychlorinated aromatic hydrocarbonas

Abstract

Aryl hydrocarbon (Ah) receptor (Ah-agonist) effects of environmental samples containing polychlorinated aromatic hydrocarbons were evaluated using a 7-ethoxyresorufin-O-deethylase (EROD) assay of a primary hepatocyte culture from grass carp (Ctenopharyngodon idellus). The results were compared with those obtained from the assay using the rat hepatoma cell line H4IIE and chemical analysis using high-resolution gas chromatography/high-resolution mass spectrometry (HRGC/HRMS). A dose-response relationship was observed between the EROD activities, either from primary hepatocyte culture assay or from H4IIE assay, and concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The results showed that the assay based on the H4IIE cell line (EC 50 =0.83 pg/mL) is more sensitive to TCDD than the assay based on primary hepatocyte culture (EC 50 =9.7 pg/mL). In tests of environmental samples, the results from the assay using primary hepatocyte culture were comparable to those from the assay using the H4IIE cell line and chemical analysis of concentrations of mixtures of polychlorinated dibenzo-p-dioxin and dibenzofuran (PCDD/PCDF). The lack of a change in the activities of glutathione-S-transferase (GST) and lactate dehydrogenase (LDH) in cell culture upon exposure to TCDD indirectly indicates that the compound is persistent to biodegradation in the cell culture system. r 2004 Elsevier Inc. All rights reserved.

Single and combined effects of selected pharmaceuticals at sublethal concentrations on multiple biomarkers in Carassius auratus

“…Acetylcholinesterase (AChE) is a biomarker for anti-cholinesterase inhibitors and could be used to indicate neurological toxicity of chemicals (Reyes et al 2011). Biotransformation phase I enzyme 7-ethoxyresorufin O-deethylase (EROD) and phase II enzymes glutathione S-transferase (GST) activities have been used as biomarkers for exposure to many groups of contaminants such as polychlorinated aromatic hydrocarbons and estrogenic pollutants (Wan et al 2004;Teles et al 2005). The antioxidant defense enzymes such as superoxide dismutase (SOD) is the first and most important line of defense and this enzyme has been proposed as a biomarker of contaminant mediated oxidative stress in a variety of freshwater and marine organisms (Orbea et al 2002).…”

Effects of astaxanthin on antioxidant capacity of golden pompano (Trachinotus ovatus) in vivo and in vitro

“…Hepatocytes were isolated according to the methods of Wan et al (2004) with some modifications. In the procedure, a fish was kept in 0.01% potassium permanganate solution for half an hour, after that, its skin was sterilized by alcohol, and its abdomen was dissected with sterile instruments from the anus toward the head.…”

Dioxin-like compounds in agricultural soils near e-waste recycling sites from Taizhou area, China: Chemical and bioanalytical characterization

“…The program is a four-parameter logistic equation (Wan et al, 2004). The bioassay-derived TCDD equivalent (expressed as TEQ bio ) for each soil sample was estimated by comparing value to the TCDD dose-response curve created using the standard solutions on the same plate.…”