Mequindox (MEQ) is a synthetic antimicrobial agent of quinoxaline-1,4-dioxide group (QdNOs). The liver is regarded as the toxicity target of QdNOs, and the role of N → O group-associated various toxicities mediated by QdNOs is well recognized. However, the mechanism underlying the in vivo effects of MEQ on the liver, and whether the metabolic pathway of MEQ is altered in response to the pathophysiological conditions still remain unclear. We now provide evidence that MEQ triggers oxidative damage in the liver. Moreover, using LC/MS-ITTOF analysis, two metabolites of MEQ were detected in the liver, which directly confirms the potential connection between N → O group reduction metabolism of MEQ and liver toxicity. The gender difference in MEQ-induced oxidative stress might be due to adrenal toxicity and the generation of M4 (2-isoethanol 1-desoxymequindox). Furthermore, up-regulation of the MAPK and Nrf2-Keap1 family and phase II detoxifying enzymes (HO-1, GCLC and NQO1) were also observed. The present study demonstrated for the first time the protein peroxidation and a proposal metabolic pathway after chronic exposure of MEQ, and illustrated that the MAPK, Nrf2-Keap1 and NF-кB signaling pathways, as well as the altered metabolism of MEQ, were involved in oxidative toxicity mediated by MEQ in vivo.

College of Life Science Yangtze University Jingzhou China

Department of Biosciences COMSATS Institute of Information Technology Sahiwal Pakistan

Department of Chemistry Faculty of Science University of Hradec Kralove Hradec Kralove Czech Republic

Hubei Collaborative Innovation Center for Animal Nutrition and Feed Safety Wuhan Hubei China

MOA Laboratory for Risk Assessment of Quality and Safety of Livestock and Poultry Products Huazhong Agricultural University Wuhan Hubei 430070 China

National Reference Laboratory of Veterinary Drug Residues and MAO Key Laboratory for Detection of Veterinary Drug Residues Huazhong Agricultural University Wuhan Hubei 430070 China

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Cheng G. et al.. Systematic and Molecular Basis of the Antibacterial Action of Quinoxaline 1,4-Di-N-Oxides against Escherichia coli. Plos one 10, e0136450 (2015). PubMed PMC

Liu Q. Y. et al.. Further investigations into the genotoxicity of quinoxaline-di-N-oxides and their primary metabolites. Food Chem Toxicol 93, 145–157 (2016). PubMed

Vicente E. et al.. Selective activity against Mycobacteriumtuberculosis of new quinoxaline 1,4-di-N-oxides. Bioorgan Med Chem 17, 385–389 (2009). PubMed

Wang X. et al.. Two generation reproduction and teratogenicity studies of feeding cyadox in Wistar rats. Food Chem Toxicol 49, 1068–1079 (2011). PubMed

Wang X. et al.. Genotoxic risk of quinocetone and its possible mechanism in in-vitro studies. Toxicol Res, doi: 10.1039/C1035TX00341E (2015). PubMed DOI PMC

Wang X. et al.. Deoxidation rates play a critical role in DNA damage mediated by important synthetic drugs, quinoxaline 1,4-dioxides. Chem Res Toxicol 28, 470–481 (2015). PubMed

Wu Y. et al.. Development of a high-performance liquid chromatography method for the simultaneous quantification of quinoxaline-2-carboxylic acid and methyl-3-quinoxaline-2-carboxylic acid in animal tissues. J Chromatogr A 1146, 1–7 (2007). PubMed

Zhang K. et al.. Investigation of quinocetone-induced mitochondrial damage and apoptosis in HepG2 cells and compared with its metabolites. Environ Toxicol Pha 39, 555–567 (2015). PubMed

Carta A., Corona P. & Loriga M. Quinoxaline 1,4-dioxide: a versatile scaffold endowed with manifold activities. Curr Med Chem 12, 2259–2272 (2005). PubMed

Chen Q. et al.. Investigation of the genotoxicity of quinocetone, carbadox and olaquindox in vitro using Vero cells. Food Chem Toxicol 47, 328–334 (2009). PubMed

Ihsan A. et al.. Acute and subchronic toxicological evaluation of Mequindox in Wistar rats. Regul Toxicol Phar 57, 307–314 (2010). PubMed

Ihsan A. et al.. Long-term mequindox treatment induced endocrine and reproductive toxicity via oxidative stress in male Wistar rats. Toxicol Appl Pharm 252, 281–288 (2011). PubMed

Ihsan A. et al.. Genotoxicity of quinocetone, cyadox and olaquindox in vitro and in vivo. Food Chem Toxicol 59, 207–214 (2013). PubMed

Huang L. et al.. Metabolism, Distribution, and Elimination of Mequindox in Pigs, Chickens, and Rats. J Agri Food Chem 63, 9839–9849 (2015). PubMed

Cheng G. et al.. Quinoxaline 1,4-di-N-Oxides: Biological Activities and Mechanisms of Actions. Front Pharmacol 7, 64 (2016). PubMed PMC

Chen Q. et al.. Characterization of carbadox-induced mutagenesis using a shuttle vector PSP189 in mammalian cells. Mutat Res 638, 11–16 (2008). PubMed

Hao L., Chen Q. & Xiao X. 2006. Molecular mechanism of mutagenesis induced by olaquindox using a shuttle vector pSP189/mammalian cell system. Mutat Res 599, 21–25 (2006). PubMed

Ganley B., Chowdhury G., Bhansali J., Daniels J. S. & Gates K. S. Redox-activated, hypoxia-selective DNA cleavage by quinoxaline 1,4-di-N-oxide. Bioorgan Med Chem 9, 2395–2401 (2001). PubMed

Zhang K. et al.. Cytotoxicity and genotoxicity of 1,4-bisdesoxyquinocetone, 3-methyl-quinoxaline-2-carboxylic acid (MQCA) in human hepatocytes. Res Vet Sci 93, 1393–1401 (2012). PubMed

Ihsan A. et al.. Genotoxicity evaluation of Mequindox in different short-term tests. Food Chem Toxicol 51, 330–336 (2013). PubMed

Wang X. et al.. Metabolites and JAK/STAT pathway were involved in the liver and spleen damage in male Wistar rats fed with mequindox. Toxicology 280, 126–134 (2011). PubMed

Azqueta A. et al.. A quinoxaline 1,4-di-N-oxide derivative induces DNA oxidative damage not attenuated by vitamin C and E treatment. Chemico-biol Interact 168, 95–105 (2007). PubMed

Chowdhury G., Kotandeniya D., Daniels J. S., Barnes C. L. & Gates K. S. Enzyme-activated, hypoxia-selective DNA damage by 3-amino-2 quinoxalinecarbonitrile 1,4-di-N-oxide. Chem Res Toxicol 17, 1399–1405 (2004). PubMed

Wang X. et al.. Fumonisins: oxidative stress-mediated toxicity and metabolism in vivo and in vitro. Arch Toxicol, doi: 10.1007/s00204-00015-01604-00208 (2015). PubMed DOI

Liu J. et al.. Mequindox induced cellular DNA damage via generation of reactive oxygen species. Mutat Res 741, 70–75 (2012). PubMed

Huang X. J. et al.. ROS mediated cytotoxicity of porcine adrenocortical cells induced by QdNOs derivatives in vitro. Chem-biol Interact 185, 227–234 (2010). PubMed

Zou J. et al.. Olaquindox-induced genotoxicity and oxidative DNA damage in human hepatoma G2 (HepG2) cells. Mutat Res 676, 27–33 (2009). PubMed

Wang X. et al.. High risk of adrenal toxicity of N1-desoxy quinoxaline 1,4-dioxide derivatives and the protection of oligomeric proanthocyanidins (OPC) in the inhibition of the expression of aldosterone synthetase in H295R cells. Toxicol 341–343, 1–16 (2016). PubMed

Huang X. J. et al.. Long-term dose-dependent response of Mequindox on aldosterone, corticosterone and five steroidogenic enzyme mRNAs in the adrenal of male rats. Toxicol Lett 191, 167–173 (2009). PubMed

Gong X., Ivanov V. N. & Hei T. K. 2,3,5,6-Tetramethylpyrazine (TMP) down-regulated arsenic-induced heme oxygenase-1 and ARS2 expression by inhibiting Nrf2, NF-kappaB, AP-1 and MAPK pathways in human proximal tubular cells. Arch Toxicol 90, 2187–200 (2015). PubMed PMC

Kim H. G. et al.. Endosulfan induces COX-2 expression via NADPH oxidase and the ROS, MAPK, and Akt pathways. Arch Toxicol 89, 2039–2050 (2015). PubMed

Liao Y. C., Chen Y. F. & Lee T. C. Increased susceptibility of H-Ras(G12V)-transformed human urothelial cells to the genotoxic effects of sodium arsenite. Arch Toxicol 89, 1971–1979 (2015). PubMed

Romanov V., Whyard T. C., Waltzer W. C., Grollman A. P. & Rosenquist T. Aristolochic acid-induced apoptosis and G2 cell cycle arrest depends on ROS generation and MAP kinases activation. Arch Toxicol 89, 47–56 (2015). PubMed

Wang X. et al.. The critical role of oxidative stress in the toxicity and metabolism of quinoxaline 1,4-di-N-oxides in vitro and in vivo. Drug Metab Rev 48, 159–182 (2016). PubMed

Watanabe T., Sekine S., Naguro I., Sekine Y. & Ichijo H. Apoptosis Signal-regulating Kinase 1 (ASK1)-p38 Pathway- dependent Cytoplasmic Translocation of the Orphan Nuclear Receptor NR4A2 Is Required for Oxidative Stress-induced Necrosis. J Biol Chem 290, 10791–10803 (2015). PubMed PMC

Ze Y. et al.. Molecular mechanism of titanium dioxide nanoparticles-induced oxidative injury in the brain of mice. Chemosphere 92, 1183–1189 (2013). PubMed

Paradies G., Petrosillo G., Pistolese M. & Ruggiero F. M. The effect of reactive oxygen species generated from the mitochondrial electron transport chain on the cytochrome c oxidase activity and on the cardiolipin content in bovine heart submitochondrial particles. FEBS lett 466, 323–326 (2000). PubMed

Kimura M. et al.. Molecular basis distinguishing the DNA binding profile of Nrf2- Maf heterodimer from that of Maf homodimer. J Biol Chem 282, 33681–33690 (2007). PubMed

Kobayashi M. & Yamamoto M. Molecular mechanisms activating the Nrf2- Keap1 pathway of antioxidant gene regulation. Antioxid Redox Sign 7, 385–394 (2005). PubMed

Li W. & Kong A. N. Molecular mechanisms of Nrf2-mediated antioxidant response. Mole Carcinogen 48, 91–104 (2009). PubMed PMC

Pedruzzi L. M., Stockler-Pinto M. B., Leite M. & Mafra D. Jr. Nrf2-keap1 system versus NF-kappaB: the good and the evil in chronic kidney disease? Biochimie 94, 2461–2466 (2012). PubMed

Lee S. E. et al.. Induction of heme oxygenase-1 inhibits cell death in crotonaldehyde-stimulated HepG2 cells via the PKC-delta-p38-Nrf2 pathway. Plos one 7, e41676 (2012). PubMed PMC

Yang C. M., Huang S. M., Liu C. L. & Hu M. L. Apo-8′-lycopenal induces expression of HO-1 and NQO-1 via the ERK/p38-Nrf2-ARE pathway in human HepG2 cells. J Agr Food Chem 60, 1576–1585 (2012). PubMed

Zhao D. X. et al.. Reactive oxygen species-dependent JNK downregulated olaquindox-induced autophagy in HepG2 cells. J Appl Toxicol 35, 709–716 (2015). PubMed

Zhao W. X. et al.. Olaquindox-induced apoptosis is suppressed through p38 MAPK and ROS-mediated JNK pathways in HepG2 cells. Cell Biol Toxicol 29, 229–238 (2013). PubMed

Yang W. et al.. Quinocetone triggers oxidative stress and induces cytotoxicity and genotoxicity in human peripheral lymphocytes of both genders. J Sci Food Agr 93, 1317–1325 (2013). PubMed

Yu M. et al.. Nrf2/ARE is the potential pathway to protect Sprague-Dawley rats against oxidative stress induced by quinocetone. Regul Toxicol Pharm 66, 279–285 (2013). PubMed

Wang X. et al.. Genomic and proteomic analysis of the inhibition of synthesis and secretion of aldosterone hormone induced by quinocetone in NCI-H295R cells. Toxicology 350–352, 1–14 (2016). PubMed

Wang X. et al.. A chronic toxicity study of cyadox in Wistar rats. Regul. Toxicol. Phar. 59, 324–333 (2011). PubMed

Wang X. et al.. Acute and sub-chronic oral toxicological evaluations of quinocetone in Wistar rats. Regul Toxicol Phar 58, 421–427 (2010). PubMed

Wang X. et al.. Mechanism of adrenocortical toxicity induced by quinocetone and its bidesoxy quinocetone metabolite in porcine adrenocortical cells in vitro. Food Chem Toxicol 84, 115–124 (2015). PubMed

Zhang K. et al.. Identification of oxidative stress and responsive genes of HepG2 cells exposed to quinocetone, and compared with its metabolites. Cell Biol Toxicol 30, 313–329 (2014). PubMed

Dai C., Tang S., Li D., Zhao K. & Xiao X. Curcumin attenuates quinocetone-induced oxidative stress and genotoxicity in human hepatocyte L02 cells. Toxicol Mech Method 25, 340–346 (2015). PubMed

Li Z. et al.. Research on olaquindox induced endoplasmic reticulum stress related apoptosis on nephrotoxicity. J Hygi Res 44, 444–450 (2015). PubMed

Zhang C. M. et al.. TNFR1/TNF-alpha and mitochondria interrelated signaling pathway mediates quinocetone-induced apoptosis in HepG2 cells. Food Chem Toxicol 62, 825–838 (2013). PubMed

Zhang T. et al.. c-Myc influences olaquindox-induced apoptosis in human hepatoma G2 cells. Mol Cell Biochem 354, 253–261 (2011). PubMed

Zou J. et al.. Olaquindox induces apoptosis through the mitochondrial pathway in HepG2 cells. Toxicol 285, 104–113 (2011). PubMed

Huang X. J. et al.. Interactions of NADPH oxidase, renin-angiotensin-aldosterone system and reactive oxygen species in mequindox-mediated aldosterone secretion in Wistar rats. Toxicol Lett 198, 112–118 (2010). PubMed

Yu M. et al.. Quinocetone-induced Nrf2/HO-1 pathway suppression aggravates hepatocyte damage of Sprague-Dawley rats. Food Chem Toxicol 69, 210–219 (2014). PubMed

Wang D. et al.. Pu-erh black tea supplementation decreases quinocetone-induced ROS generation and oxidative DNA damage in Balb/c mice. Food Chem Toxicol 49, 477–484 (2011). PubMed

Zhao X. J. et al.. Dynamic metabolic response of mice to acute mequindox exposure. J Proteome 10, Res, 5183–5190 (2011). PubMed

Liu Z. Y., Huang L. L., Chen D. M. & Yuan Z. H. Metabolism of mequindox in liver microsomes of rats, chicken and pigs. Rapid Commun Mass 24, 909–918 (2010). PubMed

Junnotula V., Sarkar U., Sinha S. & Gates K. S. Initiation of DNA strand cleavage by 1,2,4-benzotriazine 1,4-dioxide antitumor agents: mechanistic insight from studies of 3-methyl-1,2,4-benzotriazine 1,4-dioxide. J Am Chem Soc 131, 1015–1024 (2009). PubMed PMC

Poole J. S. et al.. Photochemical electron transfer reactions of tirapazamine. J Photoch Photobio 75, 339–345 (2002). PubMed

El-Khatib M., Geara F., Haddadin M. J. & Gali-Muhtasib H. Cell death by the quinoxaline dioxide DCQ in human colon cancer cells is enhanced under hypoxia and is independent of p53 and p21. Radiat Oncol, doi: 10.1186/1748-717X-5-107 (2010). PubMed DOI PMC

Yang H. Y. & Lee T. H. Antioxidant enzymes as redox-based biomarkers: a brief review. BMB Rep 48, 200–208 (2015). PubMed PMC

Shi J. et al.. Decreasing GSH and increasing ROS in chemosensitivity gliomas with IDH1 mutation. Tumour Biol 36, 655–662 (2015). PubMed

Liu Z. Y. & Sun Z. L. The metabolism of carbadox, olaquindox, mequindox, quinocetone and cyadox: an overview. J Med Chem 9, 1017–1027 (2013). PubMed

Prasad S., Ravindran J. & Aggarwal B. B. NF-kappaB and cancer: how intimate is this relationship. Mol Cell Biochem 336, 25–37 (2010). PubMed PMC

Gui S. et al.. Renal injury and Nrf2 modulation in mouse kidney following chronic exposure to TiO(2) nanoparticles. J Agri Food Chem 61, 8959–8968 (2013). PubMed

Luo L. et al.. Butylated hydroxyanisole induces distinct expression patterns of Nrf2 and detoxification enzymes in the liver and small intestine of C57BL/6 mice. Toxicol Appl Pharm 288, 339–348 (2015). PubMed

NRC. The Development of Science based Guidelines for Laboratory Animal Care, Proceedings of the November 2003 International Workshop. National Academy Press, Washington, DC (2004). PubMed

Yang Y. et al.. Olaquindox induces DNA damage via the lysosomal and mitochondrial pathway involving ROS production and p53 activation in HEK293 cells. Environ Toxicol Pha 40, 792–799 (2015). PubMed

Sánchez-Gómez F. J. et al.. Detoxifying Enzymes at the Cross-Roads of Inflammation, Oxidative Stress, and Drug Hypersensitivity: Role of Glutathione Transferase P1-1 and Aldose Reductase. Front Pharmacol 7, 237 (2016). PubMed PMC

Mequindox-Induced Kidney Toxicity Is Associated With Oxidative Stress and Apoptosis in the Mouse