title: Reactive oxygen species mediate Terbufos-induced apoptosis in mouse testicular cell lines via the modulation of cell cycle and pro-apoptotic proteins.
abstract: Terbufos (S-t-butylthiomethyl-O,O-diethyl phosphorodithioate) is a highly toxic organophosphate which is extensively used as an insecticide and nematicide. Chronic exposure to terbufos causes neuronal injury and predisposes to neurodegenerative diseases. Accumulating evidence has shown that the exposure to terbufos, as an occupational risk factor, may also cause reproductive disorders. However, the exact mechanisms of reproductive toxicity remain unclear. The present study aimed to investigate the toxic effect of terbufos on testicular cells and to explore the mechanism of toxicity on a cellular level. The cytotoxic effects of terbufos on mouse immortalized spermatogonia (GC-1), spermatocytes (GC-2), Leydig (TM3), and Sertoli (TM4) cell lines were assessed by MTT assays, caspase activation, flow cytometry, TUNEL assay, Western blot, and cell cycle analysis. The exposure to different concentrations of terbufos ranging from 50 to 800 μM for 6 h caused significant death in all the used testicular cell lines. Terbufos increased reactive oxygen species (ROS) production, reduced mitochondrial membrane potential, and initiated apoptosis, which was confirmed by a dose-dependent increase in the number of TUNEL-positive apoptotic cells. Blocking ROS production by N-acetyl cysteine (NAC) protected GC-1 cells from terbufos-induced cell death. The results demonstrated that terbufos induces ROS, apoptosis, and DNA damage in testicular cell lines and it should be considered potentially hazardous to testis. Together, this study provided potential molecular mechanisms of terbufos-induced toxicity in testicular cells and suggests a possible protective measure. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1888-1898, 2016.
authors.affiliation: Department of Biotechnology, Chia Nan University of Pharmacy and Science, Tainan, Taiwan.|Drug Discovery and Development Center, Chia Nan University of Pharmacy and Science, Tainan, Taiwan.
authors.forename: Jui-Hsiang
authors.initials: JH
authors.lastname: Hung
authors.affiliation: Department of Biotechnology, Chia Nan University of Pharmacy and Science, Tainan, Taiwan.
authors.forename: Chia-Yun
authors.initials: CY
authors.lastname: Chen
authors.affiliation: Department of Pharmacology, Faculty of Pharmacy, Beni-Suef University, Beni-Suef, 62514, Egypt.|Department of Pharmacology, College of Pharmacy, Sharjah Institute for Medical Research, University of Sharjah, Sharjah, 27272, UAE.
authors.forename: Hany A
authors.initials: HA
authors.lastname: Omar
authors.affiliation: Department of Orthopedics, National Cheng Kung University, College of Medicine, Tainan, Taiwan.
authors.forename: Kuo-Yuan
authors.initials: KY
authors.lastname: Huang
authors.affiliation: Department of Biological Sciences and Technology, National University of Tainan, Tainan, Taiwan.
authors.forename: Che-Chia
authors.initials: CC
authors.lastname: Tsao
authors.affiliation: Department of Biotechnology, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors.forename: Chien-Chih
authors.initials: CC
authors.lastname: Chiu
authors.affiliation: Department of Senior Citizen Service Management, Chia Nan University of Pharmacy and Science, Tainan, Taiwan.
authors.forename: Yi-Ling
authors.initials: YL
authors.lastname: Chen
authors.affiliation: Department of Cosmetic Application & Management, Far East University, Tainan, Taiwan.
authors.forename: Po-Han
authors.initials: PH
authors.lastname: Chen
authors.affiliation: Department of Biological Sciences and Technology, National University of Tainan, Tainan, Taiwan.
authors.forename: Yen-Ni
authors.initials: YN
authors.lastname: Teng
mesh_terms: D000818:Animals; D000969:Antinematodal Agents / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism*; D002453:Cell Cycle / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D007306:Insecticides / Q000633:toxicity*; D008297:Male; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D051379:Mice; D009946:Organothiophosphorus Compounds / Q000633:toxicity*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013737:Testis / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism
chemical_list: D000969:Antinematodal Agents; D051017:Apoptosis Regulatory Proteins; D007306:Insecticides; D009946:Organothiophosphorus Compounds; D017382:Reactive Oxygen Species; C012568:terbufos
keywords: caspase-3; reactive oxygen species; terbufos