Synergistic Activity of Ketoconazole and Miconazole with Prochloraz in Inducing Oxidative Stress, GSH Depletion, Mitochondrial Dysfunction, and Apoptosis in Mouse Sertoli TM4 Cells

• Published in: International journal of molecular sciences Vol. 23; no. 10; p. 5429
• Main Authors: Petricca, Sabrina, Celenza, Giuseppe, Luzi, Carla, Cinque, Benedetta, Lizzi, Anna Rita, Franceschini, Nicola, Festuccia, Claudio, Iorio, Roberto
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 12.05.2022; MDPI
• Subjects: Acetylcysteine; Antifungal agents; Antioxidants; Apoptosis; Azoles; Cell cycle; Cooperation; Cytotoxicity; Depletion; DNA damage; Drugs; Endocrine disruptors; Enzymes; Fertility; Fluconazole; Fungicides; Heterocyclic compounds; Imidazole; Infertility; Ketoconazole; Lipid peroxidation; Males; Miconazole; miconazole and ketoconazole; Mitochondria; mouse Sertoli TM4 cells; Oxidants; Oxidative stress; Oxidizing agents; Pesticides; Pollutants; Prochloraz; Proteins; Sertoli cells; Sperm; Spermatogenesis; Steroids; Superoxide dismutase; synergistic effects; Toxicants; Toxicity; ROS generation and apoptosis; fluconazole; prochloraz; GSH homeostasis; mouse Sertoli TM4 cells; synergistic effects; miconazole and ketoconazole; enzymatic activity; SOD; oxidative stress; mitochondrial activity
• DOI: 10.3390/ijms23105429
• ISSN: 1422-0067; 1661-6596

Triazole and imidazole fungicides represent an emerging class of pollutants with endocrine-disrupting properties. Concerning mammalian reproduction, a possible causative role of antifungal compounds in inducing toxicity has been reported, although currently, there is little evidence about potential cooperative toxic effects. Toxicant-induced oxidative stress (OS) may be an important mechanism potentially involved in male reproductive dysfunction. Thus, to clarify the molecular mechanism underlying the effects of azoles on male reproduction, the individual and combined potential of fluconazole (FCZ), prochloraz (PCZ), miconazole (MCZ), and ketoconazole (KCZ) in triggering in vitro toxicity, redox status alterations, and OS in mouse TM4 Sertoli cells (SCs) was investigated. In the present study, we demonstrate that KCZ and MCZ, alone or in synergistic combination with PCZ, strongly impair SC functions, and this event is, at least in part, ascribed to OS. In particular, azoles-induced cytotoxicity is associated with growth inhibitory effects, G0/G1 cell cycle arrest, mitochondrial dysfunction, reactive oxygen species (ROS) generation, imbalance of the superoxide dismutase (SOD) specific activity, glutathione (GSH) depletion, and apoptosis. N-acetylcysteine (NAC) inhibits ROS accumulation and rescues SCs from azole-induced apoptosis. PCZ alone exhibits only cytostatic and pro-oxidant properties, while FCZ, either individually or in combination, shows no cytotoxic effects up to 320 µM.