E r J V R a b a A R R 2 A A K F C B P 1 4 b t T u w 2 a h 2 N a p B 9 h 1 Environmental Toxicology and Pharmacology 42 (2016) 63–68 Contents lists available at ScienceDirect Environmental Toxicology and Pharmacology j o ur na l ho mepage: www.elsev ier .com/ locate /e tap xposure to fipronil elevates systolic blood pressure and disturbs elated biomarkers in plasma of rats oao Leandro Chaguria, Antonio Francisco Godinhoa, Daniel Franç a Hortaa, ictor Hugo Gonç alves-Rizzib, Jose Sergio Possomato-Vieirab, egina Aparecida Nascimentob, Carlos Alan Dias-Juniora,b,∗ Center for Toxicological Assistance, Biosciences Institute of Botucatu, Sao Paulo State University – UNESP, Botucatu, Sao Paulo, Brazil Department of Pharmacology, Biosciences Institute of Botucatu, Sao Paulo State University – UNESP, Botucatu, Sao Paulo, Brazil r t i c l e i n f o rticle history: eceived 31 October 2015 eceived in revised form 2 December 2015 ccepted 26 December 2015 vailable online 31 December 2015 eywords: ipronil a b s t r a c t Recent reports show that fipronil affects non-target organisms, including environmental species popu- lations and potentially humans. We aimed to examine if fipronil exposure affects the systolic blood pressure and related biomarkers. Thus, fipronil was orally administered to rats (30 mg/kg/day) during 15 days (Fipronil group) or physiological solution (Control group). While fipronil increased signifi- cantly the systolic blood pressure (158 ± 13 mmHg), no significant changes were observed in Control group (127 ± 3 mmHg). Significantly, higher levels of fipronil in plasma were observed in Fipronil group (0.46 ± 0.09 �g/mL versus 0.17 ± 0.11 �g/mL in Control group). Fipronil group showed lower weight gain compared with Control group. While fipronil resulted in higher concentrations of endothelin-1, reduced ardiovascular toxicity iomarkers esticides antioxidant capacity and lower levels of circulating matrix metalloproteinase 2 (MMP-2) and nitric oxide (NO) metabolites compared to Control group, no alteration was observed in serum biomarkers of renal and hepatic/biliary functional abilities. Therefore, this study suggests that fipronil causes hypertension and endothelin-1 plays a key role. Also, these findings suggest that reductions of both MMP-2 and NO may contribute with the elevation of systolic blood pressure observed with fipronil. . Introduction Fipronil [(±)-5-amino-1-(2,6-dichloro-�,�,�-trifluoro-p-tolyl)- trifluoromethylsulfinylpyrazole-3-carbonitrile] is the first mem- er of the phenylpyrazole insecticide class, and possesses a rifluoromethylsulfinyl functional group on the heterocyclic ring. he fipronil has a broad spectrum of action against insects, being sed to control fleas, ticks, termites, mole crickets, ants, root- orms, beetles, cockroaches and other insects (Tingle et al., 003). Fipronil was initially developed to replace pesticides, such s organophosphates, carbamate and pyrethroids insecticides, aving action against resistant pest strains (Narahashi et al., 007), becoming an insecticide widespread used in agriculture. arahashi et al. (2010) demonstrated that LD50 values of fipronil re 0.13 and 41 mg/kg in houseflies and rats, respectively, thus roviding highly specificity for fipronil (Narahashi et al., 2010). ∗ Corresponding author at: Department of Pharmacology, Biosciences Institute of otucatu, Sao Paulo State University (UNESP), Distrito de Rubiao Junior, S/N, 18.618- 70 Botucatu, Sao Paulo, Brazil. E-mail address: carlosjunior@ibb.unesp.br (C.A. Dias-Junior). ttp://dx.doi.org/10.1016/j.etap.2015.12.020 382-6689/© 2015 Elsevier B.V. All rights reserved. © 2015 Elsevier B.V. All rights reserved. Jackson et al. (2009) have suggested that the “no observed adverse effect level” (NOAEL) for acute oral dose of fipronil in rats is 2.5 mg/kg. Importantly, recent evidences have shown that fipronil may be a potent toxicant affecting non-target organisms, including environ- mental species populations and potentially humans (Le Faouder et al., 2007) and deleterious effects were observed on the early stages of the animal development (Udo et al., 2014; Badgujar et al., 2015; Terç ariol and Godinho, 2011). Although the fipronil’s actions are blocking the gamma-aminobutyric acid (GABA)-ergic recep- tors and glutamate (Glu)-gated chloride channels (Narahashi et al., 2007; Zhao et al., 2005), concerningly, evidence has demonstrated that fipronil induces the releasing of reactive oxygen species, result- ing in increased oxidative stress (Overmyer et al., 2007; Vidau et al., 2011). Supporting these findings, it has been shown that fipronil induced oxidative stress in kidney and brain of mice (Badgujar et al., 2015). Interestingly, we have received some cases of intox- ication with fipronil in our center for assistance and control of intoxications, who has presented symptoms typically associated with the blockade of the GABAergic receptors function (such as nausea, headache and seizures) and surprisingly, also presented elevation of blood pressure. Together, these findings point to the dx.doi.org/10.1016/j.etap.2015.12.020 http://www.sciencedirect.com/science/journal/13826689 http://www.elsevier.com/locate/etap http://crossmark.crossref.org/dialog/?doi=10.1016/j.etap.2015.12.020&domain=pdf mailto:carlosjunior@ibb.unesp.br dx.doi.org/10.1016/j.etap.2015.12.020 6 icology i fi e s m i t p w e a 2 2 t S i e u a f e fi i o 8 C T t a A t ( ( t u e B u b 2 b ( # r P o b m 2 2 e u K 4 J.L. Chaguri et al. / Environmental Tox mportance of further exploring the mechanisms responsible for pronil-induced intoxication. However, no previous study had xamined the effects of exposure to fipronil on systolic blood pres- ure, and to our knowledge, this is the first study to elucidate the echanisms involved in the very early development of fipronil- nduced hypertension in rats. Therefore, we expanded previous reports and we hypothesized hat fipronil exposure (during 15 days) increases systolic blood ressure in rats and the fipronil-induced hypertension is associated ith significant changes in circulating levels of related biomarkers: ndothelin-1, matrix metalloproteinase type 2 (MMP-2), NO and ntioxidant capacity. . Materials and methods .1. Animals and experimental design All procedures for animal experimentation were approved by he Ethics Committee, Biosciences Institute of Botucatu, Sao Paulo tate University (Protocol n◦ 620/2014), which is complied with nternational guidelines of the European Community for the use of xperimental animals. Twenty male Wistar rats (250 ± 20 g) were sed in this study. The animals were housed in standard rat cages nd maintained at 22 ◦C on a 12-h light/dark cycle, and were given ree access to water and rat chow. The animals were randomly distributed into one fipronil- xposed group (Fipronil, n = 12) and one Control group (non- pronil-exposed, Control, n = 8) for 15 days, as follows: animals n the fipronil-exposed group were treated by gavage with dose f 30 mg/kg of commercial product containing fipronil (Regent® 00WG, BASF, Agro Brazil, Sao Paulo, Brazil). The animals in the ontrol group were treated by gavage with physiological solution. he protocol of intoxication using 30 mg of fipronil via oral used in his study was chosen with basis in previous studies, which evalu- ted the dose range to fipronil (Udo et al., 2014; Moser et al., 2015). t the end of the experimental protocol (15th day), we evaluated he weight gain and the rats were anaesthetized with 2% isoflurane Isoforine, Cristalia, Sao Paulo, Brazil) at a continuous oxygen flow 2 L/min), having confirmed immobility and loss of righting reflex, he rats were submitted to euthanasia. The whole-blood samples were collected in siliconized vac- um tubes containing no additives (for serum) or lyophilized thylenediaminetetraacetic acid (EDTA) or heparin (Vacuntainer ecton-Dickinson, BD, Oxford, UK) for plasma and fractions were sed after serum separation or whole blood centrifugation, and oth were stored at −80 ◦C until use for biochemical analysis. .2. Measurement of arterial blood pressure in conscious rats Indirect systolic blood pressure (mmHg) was measured 1 h efore (1st day) and during (1st to 15th) at alternated days between 8 and 12 am) using tail-cuff plethysmography (catalog EFF 306, Insight, Ribeirao Preto, Sao Paulo, Brazil). Conscious ats were restrained for 5–10 min in a warm box (Insight, Ribeirao reto, Sao Paulo, Brazil) in a quiet room and conditioned to numer- us cuff inflation-deflation cycles by a trained operator. Systolic lood pressure was measured, and the mean of three measure- ents was recorded, as previously described (Gonç alves-Rizzi et al., 015; Nascimento et al., 2015). .3. Determination of fipronil concentrations in plasma Whole-blood levels of fipronil were determined by liquid–liquid xtraction and high-performance liquid chromatography with ltraviolet detection (HPLC-UV) system (Prominence Shimadzu®, yoto, Japan), according to the method proposed by Cid et al. and Pharmacology 42 (2016) 63–68 (2012) and adapted from Xavier et al. (2014). Briefly, blood samples were subjected to extraction by stirring with acetonitrile (Merck, Germany) and the filtered material was evaporated at room tem- perature and re-suspended with acetonitrile, then, passed with hydrophilic syringe filter with 13 mm diameter and pore 0.22 �m (PTFE membrane, VWR, Atlanta, GA, USA). A volume of 10 �L was injected into HPLC-UV, using a chromatographic column (C18). The whole-blood fipronil levels were expressed in �g/mL. 2.4. Determinations of plasma endothelin-1 and MMP-2 concentrations For measurements were used commercial enzyme immunoas- say (ELISA) kits for endothelin-1 (R&D Systems Inc, Minneapolis, MN, USA, catalog # DET100) and for MMP-2 (Abcam Inc, Cambridge, MA, USA, catalog # AB100730) plasma level assays, according to the manufacturer’s instructions. The plasma endothelin-1 and MMP-2 levels were expressed in pg/mL. 2.5. Measurements of total antioxidant capacity Serum antioxidant activity was determined by the Trolox (a water-soluble vitamin E analog, 6-hydroxy-2,5,7,8- tetramethylchroman-2-carboxylic acid, Sigma, St. Louis, MO, USA) equivalent antioxidant capacity assay as previously described (Ortner Hadžiabdić et al., 2015). Briefly, the antioxidant capacity is measured as the ability of the test compound (serum) to decrease the color of the reaction mixture by reacting directly with the ABTS (2,20-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) radical cation. The ABTS solution was mixed with 300 mL of the diluted serum sample and the reaction mixture was allowed to stand in the dark. After 3 min, the inhibition of the initial absorbance was recorded at 734 nm using an ultraviolet-visible spectrophotometer (UV 4-100, ATI Unicam, Cambridge, UK). Antioxidant activity of the sample was determined using a standard curve approach and was expressed as mmol of Trolox equivalent/L of sample. 2.6. Determinations of plasma nitrite/nitrate (NOx) concentrations The plasma NO metabolites (nitrite/nitrate) – NOx concentra- tions were determined in duplicate using the Griess reaction as described previously (Gonç alves-Rizzi et al., 2015; Nascimento et al., 2015). Briefly, 40 �L of plasma was incubated with the same volume of nitrate reductase buffer (0.1 M potassium phosphate, pH 7.5, containing 1 mM b nicotinamide adenine dinucleotide phosphate and 2 U of nitrate reductase/mL) in individual wells of a 96-well plate. Samples were allowed to incubate overnight at 37 ◦C in the dark; 8 �L of freshly prepared Griess reagent (1% sulfanilamide, 0.1% naphthylethylenediamine dihydrochloride in 5% phosphoric acid) was added to each well, and the plate was incubated for an additional 15 min at room temperature. A standard nitrate curve was obtained by incubating sodium nitrate (0.2–200 lM) with the same reductase buffer. The absorbance values were measured at 540 nm using a microplate reader (Spec- trophotometer, Synergy 4, BIOTEK, Winooski, VT, USA). The NOx levels in plasma were expressed in �mol/L. 2.7. Evaluations of liver/biliary or kidney functional abilities Serum alanine aminotransferase (ALT) and aspartate amino- transferase (AST) (IFCC method with pyridoxal phosphate activa- tion), alkaline phosphatase (ALP) (IFCC method with aminomethyl- propanol buffer), gamma-glutamyltransferase (GGT) (Szasz-Persijn method, using l-g-glutamyl-3-carboxy-4-nitroanilide), creatine kinase (CK) (N-acetylcysteine-activated IFCC method), creatinine J.L. Chaguri et al. / Environmental Toxicology and Pharmacology 42 (2016) 63–68 65 60 80 100 120 140 160 180 200 Time (days) 1st 3rd 5th 7th 9th 11th 13th 15th Control (n = 8) Fipronil (n = 12) * * * Sy st ol ic b lo od p re ss ur e (m m H g) FipronilControl 0.0 0.2 0.4 0.6 * W ho le -b lo od fi pr on il le ve ls (μ g/ m L) FipronilControl 0 20 40 60 * W ei gh t g ai n (g ) A CB Fig. 1. Systolic blood pressure (mmHg, A), whole-blood fipronil levels (�g/mL, B) and weight gain (g, C) in the non-fipronil-exposed (Control group) or fipronil-exposed ( ps were assessed by one-way analysis of variance (ANOVA) test for repeated measures. * ( k C m t t 2 w b ( w d P r ( 3 3 a c 1 F ( t e e ( F ( * Table 1 Serum levels of creatinine, creatine kinase (CK), urea, alanine aminotrans- ferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and gamma-glutamyltransferase (GGT) in the non-fipronil-exposed (Control group) or fipronil-exposed (Fipronil group) rats. Data are shown as mean ± S.E.M. Biomarkers of renal and hepatic/biliary functional abilities Control group Fipronil group Creatinine (mg/dL) 0.500 ± 0.036 0.455 ± 0.017 CK (U/L) 797 ± 143 757 ± 78 Urea (mg/dL) 39 ± 2 41 ± 2 ALT (U/L), 62 ± 3 59 ± 3 AST (U/L) 121 ± 8 123 ± 9 Fipronil group) rats. Data are shown as mean ± S.E.M. Comparisons between grou p < 0.05 versus Control group. Jaffe kinetic method) and urea (urease/glutamate dehydrogenase inetic method) were assayed on a Dimension EXL 200 Integrated hemistry System (Siemens, Erlangen, Germany), according to the anufacturer’s specifications. Total imprecision, as expressed by he coefficient of variation (CV) was less than 2.5% for all analytes ested, as previously described (Lippi et al., 2006). .8. Data analysis and statistics The results are expressed as means ± SEM. Shapiro–Wilk test as applied to verify normality of data distribution. Comparisons etween groups were assessed by one-way analysis of variance ANOVA) test for repeated measures or Student’s paired t-test ith Bonferroni’s correction post hoc test, and t-distribution and egree of freedom (df) values are presented as well. We used Graph ad Prism® 6.0 for Windows (San Diego, CA, USA) to analyze the esults. Differences were considered significant when probability *p) < 0.05. . Results .1. Effect of fipronil exposure on systolic blood pressure The effects of fipronil exposure on systolic blood pressure were ssessed in conscious rats. As shown in Fig. 1, fipronil caused signifi- ant increases in systolic blood pressure (162 ± 16, t = 4.071, df = 18; 57 ± 14, t = 4.234, df = 17; and 158 ± 13 mmHg, t = 5.327, df = 16, in ipronil group on days 11, 13 and 15 of exposition, respectively Fipronil group, *p < 0.05, Fig. 1A). No significant changes in sys- olic blood pressure were observed in Control group throughout xperimental period (Fig. 1A). The Fipronil group presented higher whole-blood fipronil lev- ls (0.46 ± 0.09 �g/mL) compared with non-fipronil-exposed rats 0.17 ± 0.11 �g/mL in Control group, t = 1.583, df = 13, *p < 0.05, ig. 1B). In addition, Fipronil group showed lower weight gain 35 ± 3 g) compared with Control group (48 ± 3 g) (t = 4.176, df = 16, p < 0.05, Fig. 1C). ALP (U/L) 27 ± 2 26 ± 1 GGT (U/L) 9 ± 0.9 9 ± 0.1 3.2. Effects of fipronil exposure in circulating biomarkers levels Fipronil group presented higher concentrations of endothelin- 1 (0.152 ± 0.008 pg/mL) compared with Control group (0.197 ± 0.019 pg/mL) (t = 1.928, df = 16, *p < 0.05, Fig. 2A). Also, lower antioxidant capacity (Throlox, 0.075 ± 0.021 versus 0.242 ± 0.080 equiv./L, t = 2.407, df = 8) and levels of circulating MMP-2 (4498 ± 300 versus 3260 ± 130 pg/mL, t = 4.072, df = 16) and NOx (13 ± 1 versus 19 ± 1 �mmol/L, t = 4.617, df = 16) were found in fipronil treated rats compared with Controls (Fipronil versus Control group, *p < 0.05, Fig. 2B–D, respectively). No changes were observed in serum levels of creatinine, creatinine kinase, urea, alanine aminotransferase, aspartate amino- transferase, alkaline phosphatase, gamma-glutamyltransferase in fipronil treated rats compared with controls (Table 1, respectively). 4. Discussion The main results of this present study reveal, for the first time, that fipronil exposure resulted in elevation of both systolic blood pressure and levels of endothelin-1. Additionally, rats exposed to fipronil presented lower weight gain and decreases in antioxidant capacity and levels of MMP-2 and NOx in plasma, and no significant changes were observed in serum levels of creatinine, creatinine 66 J.L. Chaguri et al. / Environmental Toxicology Fig. 2. Plasma endothelin-1 (A), Throlox (B), MMP-2 (C) and NOx (D) in the non fi s p k f t r M s r e h n L s pronil-exposed (Control group) or fipronil-exposed (Fipronil group) rats. Data are hown as mean ± S.E.M. Comparisons between groups were assessed by Student’s aired t-test with Bonferroni’s correction post hoc test. *p < 0.05 versus Control group. inase, urea, alanine aminotransferase, aspartate aminotrans- erase, alkaline phosphatase, gamma-glutamyltransferase. Thus, hese findings suggest that endothelin-1 may play an important ole in fipronil-induced hypertension; also, the reductions of both MP-2 and NO contributed to the elevation of systolic blood pres- ure observed with fipronil. Accidental overexposure to the fipronil by farm works has been eported since its introduction in the market in the 90s (Tingle t al., 2003). The existence of fipronil-containing foodstuffs in many ouseholds has raised concerns and the possibility of contami- ation has been considered by governments (Tingle et al., 2003; e Faouder et al., 2007; Jennings et al., 2002). Although fipronil’s electivity to the insects has been attributed to the inhibition of and Pharmacology 42 (2016) 63–68 chloride channels, which results in uncontrolled neural excita- tion and, eventually, the death of the insect (Jackson et al., 2009), lower potency has been saw by antagonizes mammalian GABAer- gic receptors (Ikeda et al., 2004). Accordingly, the human fipronil toxicity results in symptoms and complications typically associ- ated with the blockade of the GABA receptor function, featured by nausea, headache and seizures (Mohamed et al., 2004). However, adverse effects non-related to this fipronil’s mode of action has been reported in experimental studies, since low doses of fipronil revealed reproductive toxicity (Moser et al., 2015; Ohi et al., 2004), disturbed maternal behavior (Udo et al., 2014) and disruption of thyroid function in rats (Leghait et al., 2009). The present study shows that fipronil elevated the systolic blood pressure. Although the mechanisms elucidating how the fipronil exposure resulted in hypertension had never been investi- gated before, our findings show that fipronil-induced hypertension (Fig. 1) could be explained by increases of endothelin-1 and decreases of NOx levels in plasma (Fig. 2). Supporting our results, it has been suggested that the normal function of the vascular endothelial lining the circulatory system is essential for vascular homeostasis (Murakami and Simons, 2009; Koyama et al., 2013). However, if the endothelial dysfunction is present as in many vascular diseases, including hypertension, there are decreases in endothelium-dependent vasodilators (NO and prostacyclin) and increases in endothelium-dependent vasoconstrictors (endothelin- 1 and thromboxane A2) (Chiu and Chien, 2011). Accordingly, studies in humans have demonstrated that endothelin-1 levels are increased in hypertensive patients (Saito et al., 1990) and endothelin-1 receptor antagonists reduced the blood pressure in resistant hypertension (Weber et al., 2009) and improved the endothelium-dependent vasodilation in essential hypertension (Cardillo et al., 2002). In regard to the NO, it has been demonstrated that lesser NOx plasma concentrations compared with control sub- jects, confirming biologically that decreases in NO bioavailability during elevation of blood pressure (Panza et al., 1993; Kleinbongard et al., 2006). Otherwise, endothelial dysfunction has been often associated with a paradoxical increased oxidative stress, according to reduc- tion in antioxidant capacity (Ortner Hadžiabdić et al., 2015; Chiu and Chien, 2011), thus, oxidative stress induced by fipronil (Badgujar et al., 2015; Bebe and Panemangalore, 2005) may has impaired the bioavailability of NO observed here (Fig. 2). Accord- ingly, previous reports stated that acute fipronil exposure induced oxidative stress (Badgujar et al., 2015) and reduced antioxidant sta- tus (Gill and Dumka, 2013) in animals. In this concern, toxicity mediated by pesticides involves excessive production of reactive oxygen species, which may lead to alterations in the cellular antiox- idant defense system resulting in susceptibility of cells to oxidative stress (Badgujar et al., 2015; Bebe and Panemangalore, 2005; Moser et al., 2015). Reduction of cellular antioxidant status makes up the cell to become vulnerable to modify lipids, nucleic acids and proteins leading to cell disruption and increased production of lipid peroxidation induced by oxidative stress (Salminen and Paul, 2014). Thus, it is reasonable to consider that endothelial dysfunc- tion has contributed with the fipronil toxicity in the present study. In addition, decreases in MMP-2 levels in plasma were also observed with fipronil (Fig. 2), supporting earlier findings in which increases in tissue inhibitor of MMP (TIMP) were showed after fipronil exposure (Moser et al., 2015). Accordingly, both reduced levels of MMP-2 and NO could increase vessel rigidity and decrease its plasticity, contributing to increased vascular resistance and hypertension. These suggestions are consistent with previous reports that MMP-2 (a typical gelatinolytic enzyme) is decreased while the collagen deposition presents increased in arteries from hypertensive patients (Ergul et al., 2004) and from hypertension in pregnant rats (Li et al., 2014). icology c r G 1 1 e w a c e a t T a f s t ( F b t s t c I i fi i d c i a t b h ( s p 5 o d a c e t o i r b C A E J.L. Chaguri et al. / Environmental Tox Importantly, the fipronil’s mechanism is blocking the chloride hannels of GABA-ergic receptors and glutamate-gated chlo- ide channels (Narahashi et al., 2007; Zhao et al., 2005) and ABA antagonists activate vasopressin release (Isobe and Nishino, 997; Magnusson and Meyerson, 1993; Sladek and Armstrong, 987). Furthermore, vasopressin mediates the expression of the ndothelin-1 (Emori et al., 1991; Imai et al., 1992; Li et al., 2003), hich, in turn, is responsible for control the heart rate and systemic rterial pressure (Faraci, 1989). Therefore, it is possible that fipronil auses the vasopressin release and vasopressin had induced the xpression of endothelin-1, elevating the systolic blood pressure s observed in the present study. Earlier studies have reported that liver, kidney and brain are he target organs for toxic insults of fipronil (Badgujar et al., 2015; erç ariol and Godinho, 2011), owing to the fact that these tissues re extremely perfused organs and receive high blood volumes. In act, liver toxicity to fipronil has been reported in mice following ubchronic and chronic exposures (De Oliveira et al., 2012). On he other hand, this was not observed in our current short-term 15 days) study (Table 1). The present results show that although ipronil group presented lower weight gain (Fig. 1), suggesting iological toxicity, no significant alterations were observed nei- her liver/biliary or kidney functional abilities as reflected by no ignificant changes in alanine aminotransferase, aspartate amino- ransferase, alkaline phosphatase, gamma-glutamyltransferase, reatine kinase, creatinine and urea levels in plasma (Table 1). mportantly, a previous study showed that with repeated dos- ng fipronil accumulated (Moser et al., 2015) and increases in pronil levels have been related to induction of hepatic metaboliz- ng enzymes (Moser et al., 2015; Das et al., 2006). However, these ifferences could be explained, at least in part, by physiological and ompensatory responses, as well as differing levels of fipronil and ts metabolites resulted from different dosage regimen. Therefore, it should be taken into account that reference levels nd acceptable daily intakes for fipronil are set at doses much lower han those used here, which were approached to try to identify iological changes. In addition, although these doses may reflect igh peaks in exposures with high contamination or poisonings Jennings et al., 2002; Mohamed et al., 2004), a logical follow-up tudy should be driven to assess lower doses and longer exposure eriods, to start approaching environmentally relevant exposures. . Conclusion The main conclusions of the present study are that repeated ral exposition of 30 mg/kg/day of fipronil caused hypertension and isturbed related biomarkers that regulate the blood pressure. In ddition, increases in endothelin-1 and decreases in antioxidant apacity, MMP-2 and NO were observed after 15 days of fipronil xposure. However, no changes in hepatic/biliary or renal func- ion after 15 days of exposure were seen with fipronil. Therefore, ur results suggest that endothelin-1 plays a key role in fipronil- nduced hypertension and are in accordance with the notion that eductions of both MMP-2 and NO contribute to the elevation of lood pressure. onflict of interest There is no conflict of interest to declare. cknowledgements This study was funded by Fundacao de Amparo a Pesquisa do stado de Sao Paulo (FAPESP, Brazil). and Pharmacology 42 (2016) 63–68 67 References Badgujar, P.C., Pawar, N.N., Chandratre, G.A., Telang, A.G., Sharma, A.K., 2015. Fipronil induced oxidative stress in kidney and brain of mice: protective effect of vitamin E and vitamin C. Pestic. Biochem. Physiol. 118, 10–18. Bebe, F.N., Panemangalore, M., 2005. Pesticides and essential minerals modify endogenous antioxidant and cytochrome P450 in tissues of rats. J. Environ. Sci. Health B 40, 769–784. Cardillo, C., Campia, U., Kilcoyne, C.M., Bryant, M.B., Panza, J.A., 2002. Improved endothelium-dependent vasodilation after blockade of endothelin receptors in patients with essential hypertension. Circulation 105, 452–456. Chiu, J.J., Chien, S., 2011. Effects of disturbed flow on vascular endothelium: patho- physiological basis and clinical perspectives. Physiol. Rev. 91 (1), 327–387. Cid, Y.P., Ferreira, T.P., Medeiros, D.M.V.C., Oliveira, R.M., Silva, N.C.C., Scott, F.B., 2012. Determination of fipronil in bovine plasma by solid-phase extraction and liquid chromatography with ultraviolet detection. Quim. Nova 35 (10), 2063–2066. Das, P.C., Cao, Y., Cherrington, N., Hodgson, E., Rose, R.L., 2006. Fipronil induces CYP isoforms and cytotoxicity in human hepatocytes. Chem. Biol. Interact. 164 (3), 200–214. De Oliveira, P.R., Bechara, G.H., Denardi, S.E., Oliveira, R.J., Mathias, M.I., 2012. Cyto- toxicity of fipronil on mice liver cells. Microsc. Res. Tech. 75 (1), 28–35. Emori, T., Hirata, Y., Ohta, K., Kanno, K., Eguchi, S., Imai, T., Shichiri, M., Marumo, F., 1991. Cellular mechanism of endothelin-1 release by angiotensin and vaso- pressin. Hypertension 18 (2), 165–170. Ergul, A., Portik-Dobos, V., Hutchinson, J., Franco, J., Anstadt, M.P., 2004. Downreg- ulation of vascular matrix metalloproteinase inducer and activator proteins in hypertensive patients. Am. J. Hypertens. 17, 775–782. Faraci, F.M., 1989. Effects of endothelin and vasopressin on cerebral blood vessels. Am. J. Physiol. 257, H799–H803. Gill, K.K., Dumka, V.K., 2013. Antioxidant status in oral subchronic toxicity of fipronil and fluoride co-exposure in buffalo calves. Toxicol. Ind. Health 10, 1–9. Gonç alves-Rizzi, V.H., Nascimento, R.A., Possomato-Vieira, J.S., Dias-Junior, C.A., 2015. Sodium nitrite prevents both reductions in circulating nitric oxide and hypertension in 7-day lead-treated rats. Basic Clin. Pharmacol. Toxicol. Ikeda, T., Nagata, K., Kono, Y., Yeh, J.Z., Narahashi, T., 2004. Fipronil modulation of GABAA receptor single-channel currents. Pest Manag. Sci. 60, 487–492. Imai, T., Hirata, Y., Emori, T., Yanagisawa, M., Masaki, T., Marumo, F., 1992. Induc- tion of endothelin-1 gene by angiotensin and vasopressin in endothelial cells. Hypertension 19 (6), 753–757. Isobe, Y., Nishino, H., 1997. GABAergic control of Arg-vasopressin release from suprachiasmatic nucleus slice culture. Brain Res. 755 (2), 213–220. Jackson, D., Cornell, C.B., Luukinen, B., Buhl, K., Stone, D., 2009. Fipronil technical fact sheet. National Pesticide Information Center, Oregon State, pp. 1–3. Jennings, K.A., Canerdy, T.D., Keller, R.J., Atieh, B.H., Doss, R.B., Gupta, R.C., 2002. Human exposure to fipronil from dogs treated with frontline. Vet. Hum. Toxicol. 44, 301–303. Kleinbongard, P., Dejam, A., Lauer, T., Jax, T., Kerber, S., Gharini, P., Balzer, J., Zotz, R.B., Scharf, R.E., Willers, R., Schechter, A.N., Feelisch, M., Kelm, M., 2006. Plasma nitrite concentrations reflect the degree of endothelial dysfunction in humans. Free Radic. Biol. Med. 40, 295–302. Koyama, T., Ochoa-Callejero, L., Sakurai, T., Kamiyoshi, A., Ichikawa-Shindo, Y., Iinuma, N., Arai, T., Yoshizawa, T., Iesato, Y., Lei, Y., Uetake, R., Okimura, A., Yamauchi, A., Tanaka, M., Igarashi, K., Toriyama, Y., Kawate, H., Adams, R.H., Kawakami, H., Mochizuki, N., Martínez, A., Shindo, T., 2013. Vascular endothe- lial adrenomedullin-RAMP2 system is essential for vascular integrity and organ homeostasis. Circulation 127 (7), 842–853. Le Faouder, J., Bichon, E., Brunschwig, P., Landelle, R., Andre, F., Le Bizec, B., 2007. Transfer assessment of fipronil residues from feed to cow milk. Talanta 73 (4), 710–717. Leghait, J., Gayrard, V., Picard-Hagen, N., Camp, M., Perdu, E., Toutain, P.L., Viguié, C., 2009. Fipronil-induced disruption of thyroid function in rats is mediated by increased total and free thyroxine clearances concomitantly to increased activity of hepatic enzymes. Toxicology 255 (1–2), 38–44. Li, L., Galligan, J.J., Fink, G.D., Chen, A.F., 2003. Vasopressin induces vascular super- oxide via endothelin-1 in mineralocorticoid hypertension. Hypertension 41, 663–668. Li, W., Mata, K.M., Mazzuca, M.Q., Khalil, R.A., 2014. Altered matrix metalloproteinase-2 and -9 expression/activity links placental ischemia and anti-angiogenic sFlt-1 to uteroplacental and vascular remodeling and collagen deposition in hypertensive pregnancy. Biochem. Pharmacol. 89 (3), 370–385. Lippi, G., Salvagno, G.L., Montagnana, M., Brocco, G., Guidi, G.C., 2006. Influence of hemolysis on routine clinical chemistry testing. Clin. Chem. Lab. Med. 44 (3), 311–316. Magnusson, A.M., Meyerson, B.J., 1993. GABA-A agonist muscimol inhibits stimu- lated vasopressin release in the posterior pituitary of Sprague-Dawley, Wistar. Wistar-Kyoto and spontaneously hypertensive rats. Neuroendocrinology 58 (5), 519–524. Mohamed, F., Senarathna, L., Percy, A., Abeyewardene, M., Eaglesham, G., Cheng, R., Azher, S., Hittarage, A., Dissanayake, W., Sheriff, M.H., Davies, W., Buckley, N.A., Eddleston, M., 2004. Acute human self-poisoning with the N-phenylpyrazole insecticide fipronil – a GABAA-gated chloride channel blocker. J. Toxicol. Clin. Toxicol. 42, 955–963. Moser, V.C., Stewart, N., Freeborn, D.L., Crooks, J., MacMillan, D.K., Hedge, J.M., Wood, C.E., McMahen, R.L., Strynar, M.J., Herr, D.W., 2015. Assessment of serum http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0005 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0010 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0015 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0015 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0015 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0015 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0015 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0015 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0015 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0015 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0015 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0015 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0015 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0015 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0015 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0015 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0015 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0015 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0015 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0015 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0020 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0020 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0020 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0020 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0020 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0020 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0020 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0020 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0020 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0020 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0020 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0020 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0020 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0020 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0020 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0020 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0020 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0020 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0020 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0020 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0025 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0030 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0030 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0030 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0030 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0030 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0030 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0030 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0030 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0030 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0030 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0030 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0030 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0030 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0030 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0030 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0030 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0030 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0035 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0035 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0035 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0035 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0035 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0035 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0035 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0035 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0035 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0035 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0035 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0035 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0035 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0035 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0035 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0035 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0040 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0040 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0040 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0040 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0040 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0040 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0040 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0040 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0040 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0040 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0040 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0040 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0040 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0040 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0040 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0040 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0045 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0045 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0045 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0045 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0045 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0045 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0045 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0045 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0045 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0045 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0045 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0045 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0045 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0045 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0045 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0045 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0045 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0045 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0045 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0045 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0050 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0050 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0050 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0050 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0050 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0050 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0050 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0050 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0050 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0050 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0050 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0050 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0050 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0050 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0050 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0050 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0055 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0060 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0060 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0060 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0060 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0060 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0060 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0060 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0060 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0060 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0060 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0060 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0060 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0060 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0060 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0060 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0060 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0060 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0060 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0060 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0070 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0070 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0070 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0070 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0070 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0070 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0070 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0070 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0070 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0070 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0070 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0070 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0070 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0070 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0065 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0065 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0065 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0065 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0065 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0065 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0065 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0065 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0065 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0065 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0065 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0065 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0065 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0065 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0065 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0065 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0065 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0065 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0075 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0075 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0075 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0075 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0075 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0075 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0075 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0075 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0075 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0075 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0075 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0075 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0075 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0075 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0075 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0075 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0075 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0080 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0080 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0080 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0080 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0080 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0080 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0080 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0080 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0080 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0080 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0080 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0080 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0080 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0080 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0085 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0085 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0085 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0085 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0085 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0085 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0085 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0085 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0085 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0085 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0085 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0085 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0085 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0085 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0085 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0085 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0090 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0090 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0090 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0090 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0090 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0090 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0090 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0090 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0090 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0090 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0090 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0090 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0090 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0090 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0090 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0090 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0090 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0090 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0090 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0095 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0095 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0095 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0095 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0095 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0095 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0095 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0095 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0095 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0095 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0095 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0095 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0095 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0095 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0095 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0095 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0095 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0095 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0095 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0100 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0100 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0100 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0100 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0100 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0100 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0100 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0100 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0100 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0100 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0100 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0100 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0100 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0100 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0100 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0100 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0105 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0115 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0115 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0115 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0115 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0115 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0115 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0115 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0115 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0115 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0115 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0115 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0115 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0115 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0115 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0115 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0110 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0120 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0120 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0120 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0120 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0120 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0120 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0120 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0120 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0120 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0120 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0120 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0120 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0120 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0120 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0120 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0120 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0120 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0125 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0130 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 6 icology M N N N O O O P S S 8 J.L. Chaguri et al. / Environmental Tox biomarkers in rats after exposure to pesticides of different chemical classes. Toxicol. Appl. Pharmacol. 282 (2), 161–174. urakami, M., Simons, M., 2009. Regulation of vascular integrity. Mol. Med. (Berl.) 87 (6), 571–582. arahashi, T., Zhao, X., Ikeda, T., Nagata, K., Yeh, J.Z., 2007. Differential actions of insecticides on target sites: basis for selective toxicity. Hum. Exp. Toxicol. 4, 361–366. arahashi, T., Zhao, X., Ikeda, T., Salgado, V.L., Yeh, J.Z., 2010. Glutamate-activated chloride channels: unique fipronil targets present in insects but not in mammals. Pestic. Biochem. Physiol. 97 (2), 149–152. ascimento, R.A., Mendes, G., Possomato-Vieira, J.S., Gonç alves-Rizzi, V.H., Kushima, H., Delella, F.K., Dias-Junior, C.A., 2015. Metalloproteinase inhibition protects against reductions in circulating adrenomedullin during lead-induced acute hypertension. Basic Clin. Pharmacol. Toxicol. 116 (6), 508–515. hi, M., Dalsenter, P.R., Andrade, A.J., Nascimento, A.J., 2004. Reproductive adverse effects of fipronil in Wistar rats. Toxicol. Lett. 146 (2), 121–127. rtner Hadžiabdić, M., Vitali Čepo, D., Rahelić, D., Božikov, V., 2015. The effect of the mediterranean diet on serum total antioxidant capacity in obese patients: a randomized controlled trial. J. Am. Coll. Nutr. 5, 1–12. vermyer, J.P., Rouse, D.R., Avants, J.K., Garrison, A.W., Delorenzo, M.E., Chung, K.W., Key, P.B., Wilson, W.A., Black, M.C., 2007. Toxicity of fipronil and its enantiomers to marine and freshwater non-targets. J. Environ. Sci. Health B 42, 471–479. anza, J.A., Casino, P.R., Kilcoyne, C.M., Quyyumi, A.A., 1993. Role of endothelium- derived nitric oxide in the abnormal endothelium-dependent vascular relaxation of patients with essential hypertension. Circulation 87, 1468–1474. aito, Y., Nakao, K., Mukoyama, M., Imura, H., 1990. Increased plasma endothelin level in patients with essential hypertension. N. Engl. J. Med. 322 (3), 205. alminen, L.E., Paul, R.H., 2014. Oxidative stress and genetic markers of suboptimal antioxidant defense in the aging brain: a theoretical review. Rev. Neurosci. 25 (6), 805–819. and Pharmacology 42 (2016) 63–68 Sladek, C.D., Armstrong, W.E., 1987. �-Aminobutyric acid antagonists stimu- late vasopressin release from organ-cultured hypothalamo-neurohypophyseal explants. Endocrinology 120, 1576–1580. Terç ariol, P.R., Godinho, A.F., 2011. Behavioral effects of acute exposure to the insec- ticide fipronil. Pestic. Biochem. Physiol. 99, 221–225. Tingle, C.C., Rother, J.A., Dewhurst, C.F., Lauer, S., King, W.J., 2003. Fipronil: environ- mental fate, ecotoxicology, and human health concerns. Rev. Environ. Contam. Toxicol. 176, 1–66. Udo, M.S., Sandini, T.M., Reis, T.M., Bernardi, M.M., Spinosa, H.S., 2014. Prenatal expo- sure to a low fipronil dose disturbs maternal behavior and reflex development in rats. Neurotoxicol. Teratol. 45, 27–33. Vidau, C., Gonzalez-Polo, R.A., Niso-Santano, M., Gomez-Sanchez, R., Bravo-San Pedro, J.M., Pizarro-Estrella, E., Blasco, R., Brunet, J.L., Belzunces, L.P., Fuentes, J.M., 2011. Fipronil is a powerful uncoupler of oxidative phosphorylation that triggers apoptosis in human neuronal cell line SHSY5Y. Neurotoxicology 32, 935–943. Weber, M.A., Black, H., Bakris, G., Krum, H., Linas, S., Weiss, R., Linseman, J.V., Wiens, B.L., Warren, M.S., Lindholm, L.H., 2009. A selective endothelin-receptor antagonist to reduce blood pressure in patients with treatment-resistant hyper- tension: a randomised, double-blind, placebo-controlled trial. Lancet 374, 1423–1431. Xavier, G., Chandran, M., George, T., Beevi, S.N., Mathew, T.B., Paul, A., Arim- boor, R., Vijayasree, V., Pradeepkumar, G.T., Rajith, R., 2014. Persistence and effect of processing on reduction of fipronil and its metabolites in chilli pepper (Capsicum annum L.) fruits. Environ. Monit. Assess. 186 (9), 5429–5437. Zhao, X., Yeh, J.Z., Salgado, V.L., Narahashi, T., 2005. Sulfone metabolite of fipronil blocks gamma-aminobutyric acid- and glutamate-activated chloride chan- nels in mammalian and insect neurons. J. Pharmacol. Exp. Ther. 314 (1), 363–373. http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0135 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0140 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0140 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0140 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0140 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0140 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0140 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0140 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0140 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0140 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0140 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0140 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0140 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0145 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0145 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0145 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0145 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0145 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0145 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0145 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0145 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0145 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0145 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0145 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0145 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0145 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0145 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0145 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0145 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0145 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0145 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0150 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0155 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0160 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0160 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0160 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0160 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0160 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0160 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0160 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0160 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0160 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0160 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0160 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0160 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0160 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0160 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0160 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0165 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0170 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0170 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0170 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0170 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0170 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0170 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0170 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0170 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0170 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0170 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0170 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0170 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0170 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0170 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0170 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0170 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0170 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0170 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0170 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0170 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0175 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0180 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0180 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0180 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0180 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0180 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0180 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0180 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0180 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0180 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0180 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0180 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0180 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0180 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0180 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0180 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0180 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0185 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0190 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0190 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0190 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0190 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0190 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0190 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0190 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0190 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0190 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0190 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0190 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0190 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0190 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0190 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0190 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0190 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0190 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0195 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0195 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0195 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0195 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0195 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0195 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0195 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0195 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0195 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0195 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0195 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0195 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0195 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0195 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0195 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0195 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0195 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0200 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0200 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0200 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0200 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0200 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0200 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0200 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0200 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0200 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0200 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0200 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0200 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0200 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0200 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0200 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0200 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0200 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0205 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0210 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0215 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0220 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 http://refhub.elsevier.com/S1382-6689(15)30159-9/sbref0225 Exposure to fipronil elevates systolic blood pressure and disturbs related biomarkers in plasma of rats 1 Introduction 2 Materials and methods 2.1 Animals and experimental design 2.2 Measurement of arterial blood pressure in conscious rats 2.3 Determination of fipronil concentrations in plasma 2.4 Determinations of plasma endothelin-1 and MMP-2 concentrations 2.5 Measurements of total antioxidant capacity 2.6 Determinations of plasma nitrite/nitrate (NOx) concentrations 2.7 Evaluations of liver/biliary or kidney functional abilities 2.8 Data analysis and statistics 3 Results 3.1 Effect of fipronil exposure on systolic blood pressure 3.2 Effects of fipronil exposure in circulating biomarkers levels 4 Discussion 5 Conclusion Conflict of interest Acknowledgements References