title: Mutagenicity and DNA-damaging potential of clenbuterol and its metabolite 4-amino-3,5-dichlorobenzoic acid in vitro.
abstract: The aim of this study was to evaluate in vitro toxicity of clenbuterol and its metabolite 4-amino-3,5-dichlorobenzoic acid. Cytotoxicity and pro-oxidative effect of both compounds were studied on human colon adenocarcinoma cell line SW 480. No significant cytotoxic effect of either compound was observed. Results of an Ames test on Salmonella typhimurium did not indicate mutagenic activity of clenbuterol on TA 98 and TA 100 strains, regardless of metabolic activation. Potential mutagenic effects of the highest clenbuterol concentration (2500 ng/ml) were observed on the TA 1535 strain. The obtained results of alkaline comet assay on isolated human lymphocytes suggested that both compounds induced an increase of primary DNA damage in a concentration-dependent manner. 4-ADBA was a slightly more potent inducer of primary DNA damage as compared to clenbuterol. Chromosomal aberration analysis showed that clenbuterol caused a statistically significant increase in the total number of aberrant cells only at the highest concentration tested (3% vs. 0.7% in the negative control). The results of this study might represent a solid frame for designing and planning future studies with both compounds, which should further clarify their mechanisms of action and genotoxic/cytogenetic effects relevant for human risk assessment.
authors.affiliation: Laboratory for Analytical Chemistry, Croatian Veterinary Institute, Savska 143, HR-10000 Zagreb, Croatia. Electronic address: vulic@veinst.hr.
authors.forename: Ana
authors.initials: A
authors.lastname: Vulić
authors.affiliation: Faculty of Food Technology and Biotechnology, Pierottijeva 6, HR-10000 Zagreb, Croatia.
authors.forename: Ksenija
authors.initials: K
authors.lastname: Durgo
authors.affiliation: Laboratory for Analytical Chemistry, Croatian Veterinary Institute, Savska 143, HR-10000 Zagreb, Croatia.
authors.forename: Jelka
authors.initials: J
authors.lastname: Pleadin
authors.affiliation: Faculty of Food Technology and Biotechnology, Pierottijeva 6, HR-10000 Zagreb, Croatia.
authors.forename: Luka
authors.initials: L
authors.lastname: Herceg
authors.affiliation: Institute for Medical Research and Occupational Health, Ksaverska cesta 2, HR-10000 Zagreb, Croatia.
authors.forename: Nevenka
authors.initials: N
authors.lastname: Kopjar
mesh_terms: D045744:Cell Line, Tumor; D002869:Chromosome Aberrations / Q000187:drug effects; D002976:Clenbuterol / Q000633:toxicity*; D020552:Comet Assay; D004249:DNA Damage / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008214:Lymphocytes / Q000187:drug effects; D009152:Mutagenicity Tests / Q000379:methods*; D012486:Salmonella typhimurium / Q000187:drug effects; D013329:Structure-Activity Relationship; D062366:para-Aminobenzoates / Q000633:toxicity*
chemical_list: D062366:para-Aminobenzoates; C000603946:4-amino-3,5-dichlorobenzoic acid; D002976:Clenbuterol
keywords: Cytotoxicity; Genotoxicity; In vitro testing; Mutagenicity; Pro-oxidative effect; ß(2)-adrenoreceptor agonist