Benomyl, a benzimidazole fungicide, induces oxidative stress and apoptosis in neural cells

Mehtap Kara; Ezgi Oztas; Rabia Ramazanoğulları; Demetrios Kouretas; Charitini Nepka; Aristides M Tsatsakis; Aristidis S Veskoukis

doi:10.1016/j.toxrep.2020.04.001

Benomyl, a benzimidazole fungicide, induces oxidative stress and apoptosis in neural cells

Toxicol Rep. 2020 Apr 11:7:501-509. doi: 10.1016/j.toxrep.2020.04.001. eCollection 2020.

Authors

Mehtap Kara¹, Ezgi Oztas¹, Rabia Ramazanoğulları¹, Demetrios Kouretas², Charitini Nepka³, Aristides M Tsatsakis⁴, Aristidis S Veskoukis²

Affiliations

¹ Faculty of Pharmacy, Department of Pharmaceutical Toxicology, Istanbul University, 34116, Istanbul, Turkey.
² Department of Biochemistry and Biotechnology, University of Thessaly, Viopolis, Mezourlo, 41500, Larissa, Greece.
³ Department of Pathology, University Hospital of Larissa, 41110, Larissa, Greece.
⁴ Center of Toxicology Science and Research, Medical School, University of Crete, Heraklion 71003, Greece.

Abstract

Fungicides are used in the agricultural sector against the harmful action of fungi, however they are potential toxic agents for the environment and the living organisms. Benomyl is a widely encountered benzimidazole fungicide that exerts its toxicity via inhibiting microtubule formation in the nervous system and the male reproductive and endocrine systems, whilst it is a known teratogen. Since toxic effects of benomyl and its molecular mechanisms are not fully understood, we aimed to detect its neurotoxic potential via evaluating cytotoxicity, oxidative stress and apoptosis in SH-SY5Y cell line. The cells were incubated with benomyl in a concentration range between 1 and 6 μM for 24 h. Our results indicated a concentration-dependent enhancement of reactive oxygen species measured through flow cytometry and DNA damage evaluated via the comet assay. Additionally, it induced apoptosis in all tested concentrations. According to the findings of the present study, benomyl is a xenobiotic, which it appears to exert its toxic action via a redox-related mechanism that, finally, induces cell apoptosis and death. We believe that this study will offer further insight in the toxicity mechanism of benomyl, although further studies are recommended in order to elucidate these mechanisms in the molecular level.

Keywords: ATCC, American Type Culture Collection; Apoptosis; BSA, Bovine serum albumin; Benomyl; DMEM-F12, Dulbecco’s modified Eagle medium: Nutrient Mixture F-12 (Ham`s); DNA, Deoxyribonucleic acid; DTNB, 55′-dithiobis-2-nitrobenzoic acid; FBS, Fetal bovine serum; GSH, Glutathione; H2DCF-DA, 2′7′-dichlorodihydrofluorescein diacetate; MFI, Median fluorescence intensity; MTT, 3-45-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide; Neural SH-SY5Y cell line; OD, Optical density; Oxidative stress; PBS, Phosphate buffered saline; PI, Propidium iodide; ROS, Reactive oxygen species; SD, Standard deviation.