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The physiological conditions of an organism may influence its ability to cope with environmental stressors, such as contaminants. Biotransformation and the endocrine system interact with each other to promote animal’s fitness. However, little is known regarding the interaction between hormones and response to pollutants such as polycyclic aromatic hydrocarbons (PAHs). In this <i>in vitro</i> study, we aimed to increase the knowledge regarding the effects of steroid hormones on ethoxyresorufin-O-deethylase (EROD) activity inhibited by contaminants. The effects on <i>in vivo</i> induced EROD activity of <i>Anguilla anguilla</i> were assessed by conducting single and combined exposures to fluoranthene (FL) and to physiological levels of two major steroid hormones (cortisol and 17ß-estradiol). Hepatic microsome exposure to the lowest concentrations of FL (0.1 and 0.3 µM), as well as to cortisol and 17ß-estradiol (E₂), led to significant EROD activity induction. However, the highest tested concentrations of FL (0.9 and 2.7 µM) significantly inhibited this enzymatic activity. When microsomes were simultaneously exposed to 0.9 µM FL and one of the hormones, both cortisol and E₂ were able to decrease the inhibitory effects, with the former completely reverting EROD activity inhibition. These findings support the idea that cortisol and E₂ can help prevent the inhibitory effects of PAHs over biotransformation enzymes, highlighting the physiological relevance of these hormones.