Effects of human pharmaceuticals on cytotoxicity, EROD activity and ROS production in fish hepatocytes

• Published in: Toxicology (Amsterdam) Vol. 196; no. 1; pp. 41 - 55
• Main Authors: Laville, N, Aı̈t-Aı̈ssa, S, Gomez, E, Casellas, C, Porcher, J.M
• Format: Journal Article
• Language: English
• Published: Shannon Elsevier Ireland Ltd 01.03.2004; Amsterdam Elsevier Science; Elsevier
• Subjects: Animals; Biological and medical sciences; Cell Line; Cell Separation; Cell Survival; Cell Survival - drug effects; Cells, Cultured; Cytochrome P-450 CYP1A1; Cytochrome P-450 CYP1A1 - metabolism; Cytotoxicity; EROD; Hepatocytes; Hepatocytes - drug effects; Hepatocytes - enzymology; Humans; Life Sciences; Medical sciences; Oncorhynchus mykiss; Oncorhynchus mykiss - metabolism; Oxidative stress; oxygen radicals; Pharmaceutical Preparations; Pharmaceuticals; Pisces; PLHC-1; Primary rainbow trout hepatocytes; Reactive Oxygen Species; Reactive Oxygen Species - metabolism; Tetrazolium Salts; Thiazoles; Toxicology; Water Pollutants, Chemical; Water Pollutants, Chemical - toxicity; Oxidative stress; Cytotoxicity; EROD; Primary rainbow trout hepatocytes; Pharmaceuticals; PLHC-1; Human; Edible fish; Reactive oxygen species; Hepatocyte; Primary; Production; Food
• ISSN: 0300-483X; 1879-3185
• DOI: 10.1016/j.tox.2003.11.002

Pharmaceuticals are found in the aquatic environment but their potential effects on non-target species like fish remain unknown. This in vitro study is a first approach in the toxicity assessment of human drugs on fish. Nine pharmaceuticals were tested on two fish hepatocyte models: primary cultures of rainbow trout hepatocytes (PRTH) and PLHC-1 fish cell line. Cell viability, interaction with cytochrome P450 1A (CYP1A) enzyme and oxidative stress were assessed by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrasodium bromide tetrazolium (MTT), 7-ethoxyresorufin- o-deethylase (EROD) and dichlorofluorescein (DCFH-DA) assays, respectively. The tested drugs were clofibrate (CF), fenofibrate (FF), carbamazepine (CBZ), fluoxetine (FX), diclofenac (DiCF), propranolol (POH), sulfamethoxazole (SFX), amoxicillin (AMX) and gadolinium chloride (GdCl 3). All substances were cytotoxic, except AMX at concentration up to 500 μM. The calculated MTT EC 50 values ranged from 2 μM (CF) to 651 μM (CBZ) in PLHC-1, and from 53 μM (FF) to 962 μM (GdCl 3) in PRTH. CF, FF, and FX were the most cytotoxic drugs and induced oxidative stress before being cytotoxic. Compared to hepatocytes from human and dog, fish hepatocytes seemed to be more susceptible to the peroxisome proliferators (PPs) CF and FF. In PLHC-1 cells none of the tested drugs induced the EROD activity whereas POH appeared as a weak EROD inducer in PRTH. Moreover, in PRTH, SFX, DiCF, CBZ and to a lesser extend, FF and CF inhibited the basal EROD activity at clearly sublethal concentrations which may be of concern at the biological and chemical levels in a multipollution context.