Protective effects of chlorogenic acid in 3-nitropropionic acid induced toxicity and genotoxicity Norberto Alarc  on-Herrera a , Saúl Flores-Maya a , Bel  en Bellido b , Ana M. García-Bores c , Ernesto Mendoza b , Guillermo  Avila-Acevedo c , Elizabeth Hern  andez-Echeagaray b, * a Laboratorio de Recursos Naturales, UBIPRO, FES-Iztacala, Universidad Nacional Aut onoma de M exico, M exico b Laboratorio de Neurofisiología del desarrollo y la neurodegeneraci on UBIMED, FES-Iztacala, Universidad Nacional Aut onoma de M exico, M exico c Laboratorio de Fitoquímica, UBIPRO, FES-Iztacala, Universidad Nacional Aut onoma de M exico, M exico article info Article history: Received 23 March 2017 Received in revised form 29 April 2017 Accepted 30 April 2017 Available online 3 May 2017 Keywords: Mitochondria Antioxidants Polyphenols Caffeoyl Neurotoxins Micronuclei abstract Mitochondrial inhibition with the toxin 3-Nitropropionic acid (3-NP) has been used to study the un- derlying mechanisms in striatal neurodegeneration, but few experiments have evaluated its toxicity and genotoxicity of in vivo administration. Furthermore, different antioxidant molecules may prevent degeneration induced by the toxic effects of 3-NP. Therefore, the purpose of this study was to evaluate the toxicity and genotoxicity induced by 3-NP (15 mg/kg) in the micronuclei assay method; also, we assessed chlorogenic acid (CGA, 100 mg/kg) for its anti-toxic and anti-genotoxic effect in damage pro- duced by in vivo treatment with 3-NP. 3-NP induced toxicity and genotoxicity. CGA administered as a co- treatment with 3-NP (3-NP þ CA) reduced toxicity by 32.76%, as a pre-treatment for 5 days only, followed by 3-NP treatment (P/CA, 3-NP) inhibiting toxicity by 24.04%, or as a pre-treatment, plus a co-treatment with 3-NP (P/CA, 3-NP þ CA) avoided any toxic effect. CGA alone did not exhibit any toxic effect. Only P/ CGA, 3-NP þ CGA group, avoided toxicity and genotoxicity, suggesting that CGA could be suitable to prevent, reduce or delay toxicity and cell death. © 2017 Elsevier Ltd. All rights reserved. 1. Introduction Neurodegenerative diseases are associated with the presence of oxidative stress which leads to neuronal death. Several experi- mental manoeuvres have been made to mimic neural degeneration by inducing oxidative stress; one example is the use of toxic mol- ecules such as the metabolite of nitropropanol, 3-nitropropionic acid (3-NP). 3-NP (C 3 H 5 NO 4, Fig. 1) is carboxylic acid with a satu- rated three carbons and a nitro group in the third carbon which gives the 3-NP its electro-activity, acid nature and specific nuclear magnetic resonance (Reynolds and Lin, 2000). 3-NP is a found in nature as a fungal toxin (Artrhirium), or in leguminous species (Indogofera and Astragalus). The accidental ingestion of 3-NP by animals or humans generate intoxication and motor function al- terations (Gould and Gustine, 1982; Gould et al., 1985; He et al., 1990, 1995) because 3-NP targets succinic dehydrogenase (SDH) due to its similarity in chemical structure to succinate (C 4 H 4 O 4 ) the endogenous substrate (Reynolds and Lin, 2000). SDH oxidizes succinate to fumarate in the Krebs cycle, and couples the oxidation of succinate to the reduction of ubiquinona. The irreversible block of SDH by 3-NP, produces reactive oxygen species (ROS), mito- chondrial dysfunction and mDNA damage, which leads to ATP depletion (Alston et al., 1977; Dr€ ose, 2013), and results in selective striatal neuronal loss. That is why this toxin has been widely used to study underlying mechanisms of Huntington Disease (HD) (Palfi et al., 1996; Brouillet et al., 2005, Hern andez-Echeagaray et al., 2012; Bhat et al., 2015). Phenolic compounds (Fig. 1) in plants are supposed to possess antioxidant, anticancer and antimicrobial properties (Balasundram et al., 2006). Chlorogenic acid (CGA), a conjugate of caffeic acid and quinic acid obtained from coffee, tea, fruits and vegetables (Clifford, 1999), has been tested under a wide concentration range as an effective antioxidant in lipid oxidation assay which is known is one Abbreviations: ANOVA, Analysis of Variance; CGA, Chlorogenic acid; HD, Hun- tington Disease; i.p., Intraperitoneal; MNC, Micronuclei cells; 3-NP, 3- nitropropionic acid; NC, Normochromatic; PC, Polychromatic; ROS, Reactive Oxy- gen Species; SDH, Succinic Dehydrogenase. * Corresponding author. Laboratorio de Neurofisiología del desarrollo y la neu- rodegeneraci on Unidad de Investigaci on en Biomedicina, FES-Iztacala Av de los Barrios #1, Los Reyes Iztacala, M exico. E-mail addresses: aehe67@gmail.com, elihernandez@unam.mx (E. Hern andez- Echeagaray). Contents lists available at ScienceDirect Food and Chemical Toxicology journal homepage: www.elsevier.com/locate/foodchemtox http://dx.doi.org/10.1016/j.fct.2017.04.048 0278-6915/© 2017 Elsevier Ltd. All rights reserved. Food and Chemical Toxicology 109 (2017) 1018e1025