RESUMEN
Anthropogenic pollution poses a threat to marine conservation by causing chronic toxic effects. Seabirds have contact throughout their lives with pollutants like plastic, metals, polychlorinated biphenyls (PCBs), and organochlorine pesticides such as hexachlorocyclohexanes (HCHs). We assessed 155 Manx shearwaters (Puffinus puffinus) stranded along the Brazilian coast, analyzing associations between organic pollutants, plastic ingestion, biomarkers (transcript levels of aryl hydrocarbon receptor, cytochrome P450-1A-5 [CYP1A5], UDP-glucuronosyl-transferase [UGT1], estrogen receptor alpha-1 [ESR1], and heat shock protein-70 genes) and enzymes activity (ethoxy-resorufin O-deethylase and glutathione S-transferase [GST]). Plastic debris was found in 29 % of the birds. The transcription of UGT1 and CYP1A5 was significantly associated with hexachlorobenzene (HCB) and PCBs levels. ESR1 was associated with HCB and Mirex, and GST was associated with Drins and Mirex. While organic pollutants affected shearwaters more than plastic ingestion, reducing plastic availability remains relevant as xenobiotics are also potentially adsorbed onto plastics.
Asunto(s)
Biomarcadores , Monitoreo del Ambiente , Bifenilos Policlorados , Contaminantes Químicos del Agua , Animales , Biomarcadores/metabolismo , Contaminantes Químicos del Agua/toxicidad , Aves , Glutatión Transferasa/metabolismo , Brasil , Plásticos , Citocromo P-450 CYP1A1/metabolismo , Citocromo P-450 CYP1A1/genética , Plaguicidas/toxicidad , Glucuronosiltransferasa/metabolismo , Glucuronosiltransferasa/genética , Receptores de Hidrocarburo de Aril/metabolismoRESUMEN
The prevalence of fragrances in various hygiene products contributes to their sensorial allure. However, fragrances can induce sensitization in the skin or respiratory system, and the mechanisms involved in this process are incompletely understood. This study investigated the intricate mechanisms underlying the fragrance's effects on sensitization response, focusing on the interplay between CYP450 enzymes, a class of drug-metabolizing enzymes, and the adaptive immune system. Specifically, we assessed the expression of CYP450 enzymes and cytokine profiles in culture of BEAS-2B and mature dendritic cells (mDC) alone or in co-culture stimulated with 2 mM of a common fragrance, cinnamyl alcohol (CA) for 20 h. CYP1A1, CYP1A2, CYP1B1, CYP2A6, and CYP2A13 were analyzed by RT-PCR and IL-10, IL-12p70, IL-18, IL-33, and thymic stromal lymphopoietin (TSLP) by Cytometric Bead Array (CBA). Through RT-PCR analysis, we observed that CA increased CYP1A2 and CYP1B1 expression in BEAS-2B, with a further increased in BEAS-2B-mDC co-culture. Additionally, exposure to CA increased IL-12p70 levels in mDC rather than in BEAS-2B-mDC co-culture. In regards to IL-18, level was higher in BEAS-2B than in BEAS-2B-mDC co-culture. A positive correlation between the levels of IL-10 and CYP1B1 was found in mDC-CA-exposed and between IL-12p70 and CYP1A1 was found in BEAS-2B after CA exposure. However, IL-12p70 and CYP1A2 as well as IL-18, IL-33, and CYP1A1 levels were negative, correlated mainly in co-culture control. These correlations highlight potential immunomodulatory interactions and complex regulatory relationships. Overall, exposure to CA enhances CYP450 expression, suggesting that CA can influence immune responses by degrading ligands on xenosensitive transcription factors.
Asunto(s)
Técnicas de Cocultivo , Sistema Enzimático del Citocromo P-450 , Citocinas , Células Dendríticas , Propanoles , Humanos , Citocinas/metabolismo , Células Dendríticas/efectos de los fármacos , Células Dendríticas/metabolismo , Sistema Enzimático del Citocromo P-450/metabolismo , Sistema Enzimático del Citocromo P-450/genética , Propanoles/toxicidad , Propanoles/metabolismo , Línea Celular , Citocromo P-450 CYP1A1/metabolismo , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1B1/genética , Citocromo P-450 CYP1B1/metabolismo , Perfumes/toxicidad , Receptores de Hidrocarburo de Aril/metabolismo , Receptores de Hidrocarburo de Aril/genética , Citocromo P-450 CYP1A2/metabolismo , Citocromo P-450 CYP1A2/genéticaRESUMEN
The aim of this study was twofold: (1) evaluate the effect of benzo[a]pyrene (BaP) on expression levels of AQP3 and Notch1 genes in HaCaT cells exposed "in vitro" and (2) investigate the possible biological role of assessed genes by bioinformatics methods. Cells were exposed to increasing concentrations of BaP (0.0-4.0 µM) for 1-4 days. After treatments, cell viability and expression levels of AhR, CYP1A1, AQP3, and Notch1 genes were evaluated. The possible biological role of assessed genes was evaluated using bioinformatics tools. Low cytotoxicity in HaCaT cells dosed with BaP was detected. A significant overexpression (p < .05) of CYP1A1, AQP3, and Notch1 was found in exposed HaCaT cells. The gene expression upregulation was dependent on AhR activation. The bioinformatics analysis showed that these genes were enriched in related cancer signaling pathways. The findings suggest that AQP3 and Notch1 are upregulated by AhR activation in HaCaT cells exposed to BaP.
Asunto(s)
Benzo(a)pireno , Citocromo P-450 CYP1A1 , Humanos , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , Benzo(a)pireno/metabolismo , Células HaCaT , Receptores de Hidrocarburo de Aril/genética , Receptores de Hidrocarburo de Aril/metabolismo , Regulación hacia ArribaRESUMEN
SUMMARY: Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that is highly expressed in various types of cancers including breast cancer. However, the role of AhR with its endogenous ligand 2-(1'H-indole-3'-carbonyl)-thiazole-4-carboxylic acid methyl ester (ITE) on the progression of breast cancer remains poorly understood. We aimed to investigate cell proliferation and migration states in breast cancer after activating AhR with the endogenous ligand ITE. Breast cancer tissue was evaluated by cell lines, immunohistochemistry, reverse transcription-polymerase chain reaction, cell proliferation, flow cytometry, migration assays and western blot techniques. We found that AhR was widely expressed in breast cancer tissues and metastasis lymph node tissues, but not in normal tissues. The expression AhR was independent between the age, grades and TNM classifications for breast cancer tissues. ITE treatment significantly induced the activation of AhR in a time-dependent manner in both MCF-7 and T47D breast cancer cell lines. Meanwhile, ITE did not affect the cell migration but significantly suppressed the cell proliferation in estrogen receptor positive (ER+) MCF-7 andT47D cells, which probably attribute to the induction of cell cycle arrest in G1 phase and shortened S phase. Further mechanism study showed that ERK1/2 and AKT signaling were required for the activation of AhR in MCF-7 cells. These data suggest that AhR is a potential new target for treating patients with breast cancer. ITE may be more potentially used for therapeutic intervention for breast cancer with the kind of ER(+).
El receptor de hidrocarburo de arilo (AhR) es un factor de transcripción activado por ligando que se expresa en gran medida en varios tipos de cáncer, incluido el cáncer de mama. Sin embargo, el papel de AhR con su ligando endógeno 2- (1'H-indol-3'-carbonil)-tiazol-4-ácido carboxílico metil éster (ITE) en la progresión del cáncer de mama sigue siendo poco conocido. Nuestro objetivo fue investigar la proliferación celular y los estados de migración en el cáncer de mama después de activar AhR con el ligando endógeno ITE. El tejido de cáncer de mama se evaluó mediante líneas celulares, inmunohistoquímica, reacción en cadena de la polimerasa con transcriptasa inversa, proliferación celular, citometría de flujo, ensayos de migración y técnicas de transferencia Western. Descubrimos que AhR se expresó ampliamente en tejidos de cáncer de mama y en linfonodos con metástasis, pero no en tejidos normales. La expresión AhR fue independiente entre la edad, grados y clasificaciones TNM para tejidos de cáncer de mama. El tratamiento con ITE indujo significativamente la activación de AhR de manera dependiente del tiempo en las líneas celulares de cancer de mama MCF-7 y T47D. Mientras tanto, ITE no afectó la migración celular, pero suprimió significativamente la proliferación celular en células MCF-7 y T47D con receptor de estrógeno positivo (ER+), lo que probablemente se atribuye a la inducción de la detención del ciclo celular en la fase G1 y la fase S acortada. Un estudio adicional del mecanismo mostró que las señales de ERK1/2 y AKT eran necesarias para la activación de AhR en las células MCF-7. Estos datos sugieren que AhR es un nuevo objetivo potencial para el tratamiento de pacientes con cáncer de mama. ITE puede ser utilizado más potencialmente en la intervención terapéutica para el cáncer de mama con el tipo de ER (+).
Asunto(s)
Humanos , Femenino , Tiazoles/administración & dosificación , Neoplasias de la Mama/patología , Receptores de Hidrocarburo de Aril/efectos de los fármacos , Indoles/administración & dosificación , Tiazoles/farmacología , Inmunohistoquímica , Receptores de Estrógenos , Western Blotting , Citocromo P-450 CYP1A1/genética , Línea Celular Tumoral/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Ensayos de Migración Celular , Citocromo P-450 CYP1B1/genética , Citometría de Flujo , Indoles/farmacologíaRESUMEN
The soluble fraction of polysaccharides from cabernet franc red wine (SFP) previously showed antitumoral effects by modulating the immune system. The present study tested the hypothesis that the SFP can regulate CYPs in vitro in HepG2 cells and in vivo in Walker-256 tumor-bearing rats. The SFP was used in the following protocols: (i) solid tumor, (ii) liquid tumor, and (iii) chemopreventive solid tumor. The SFP reduced solid tumor growth in both solid tumor protocols but did not inhibit liquid tumor development. The SFP reduced total CYP levels in the solid and liquid tumor protocols and reduced the gene expression of Cyp1a1 and Cyp2e1 in rats and CYP1A2 in HepG2 cells. An increase of N-acetylglucosaminidase activity was observed in all SFP-treated rats, and TNF-α levels increased in the solid tumor protocol in the vehicle, SFP, and vincristine (positive control) groups. The chemopreventive solid tumor protocol did not modify CYP levels in the liver or intestine or N-acetylglucosaminidase and myeloperoxidase activity in the liver. The in vitro digestion and nuclear magnetic resonance analyses suggested that SFP was minimally modified in the gastrointestinal system. In conclusion, SFP inhibited CYPs both in vivo and in vitro, likely as a result of its immunoinflammatory actions.
Asunto(s)
Vino , Ratas , Animales , Acetilglucosaminidasa , Sistema Enzimático del Citocromo P-450/metabolismo , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , Polisacáridos/farmacologíaRESUMEN
The organophosphorus pesticide chlorpyrifos, detected in water and food worldwide, has also been found in the Río Negro and Neuquén Valley, North Patagonia, Argentina, where the rainbow trout, Oncorhynchus mykiss, is one of the most abundant fish species. We analyzed whether chlorpyrifos affects the transport activity of the ATP-binding cassette protein transporters from the subfamily C (ABCC), which are critical components of multixenobiotic resistance. We exposed ex vivo O. mykiss middle intestine strips (non-polarized) and segments (polarized) for one hour to 0 (solvent control), 3, 10, and 20 µg L-1 and to 0, 10, and 20 µg L-1 chlorpyrifos, respectively. We estimated the Abcc-mediated transport rate by measuring the transport rate of the specific Abcc substrate 2,4-dinitrophenyl-S-glutathione (DNP-SG). In addition, we measured the enzymatic activity of cholinesterase, carboxylesterase, glutathione-S-transferase, and 7-ethoxyresorufin-O-deethylase (EROD, indicative of the activity of cytochrome P450 monooxygenase 1A, CYP1A). We also measured lipid peroxidation using the thiobarbituric acid reactive substances method and the gene expression of Abcc2 and genes of the AhR pathway, AhR, ARNT, and cyp1a, by qRT-PCR. Chlorpyrifos induced the DNP-SG transport rate in middle intestine strips in a concentration-dependent manner (49-71%). In polarized preparations, the induction of the DNP-SG transport rate was observed only in everted segments exposed to 20 µg L-1 chlorpyrifos (40%), indicating that CPF only stimulated the apical (luminal) transport flux. Exposure to chlorpyrifos increased GST activity by 42% in intestine strips and inhibited EROD activity (47.5%). In addition, chlorpyrifos exposure inhibited cholinesterase (34-55%) and carboxylesterase (33-42.5%) activities at all the concentrations assayed and increased TBARS levels in a concentration-dependent manner (71-123%). Exposure to 20 µgL-1 chlorpyrifos did not affect the mRNA expression of the studied genes. The lack of inhibition of DNP-SG transport suggests that chlorpyrifos is not an Abcc substrate. Instead, CPF induces the activity of Abcc proteins in the apical membrane of enterocytes, likely through a post-translational pathway.
Asunto(s)
Cloropirifos , Oncorhynchus mykiss , Plaguicidas , Contaminantes Químicos del Agua , Transportadoras de Casetes de Unión a ATP , Adenosina Trifosfato/metabolismo , Animales , Hidrolasas de Éster Carboxílico/metabolismo , Cloropirifos/farmacología , Colinesterasas , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , Sistema Enzimático del Citocromo P-450/metabolismo , Glutatión/metabolismo , Glutatión Transferasa/genética , Glutatión Transferasa/metabolismo , Intestinos , Oncorhynchus mykiss/genética , Oncorhynchus mykiss/metabolismo , Compuestos Organofosforados/metabolismo , Plaguicidas/metabolismo , ARN Mensajero/metabolismo , Solventes , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismo , Agua/metabolismo , Contaminantes Químicos del Agua/toxicidadRESUMEN
This study aims to assess breast cancer (BC) association with metals and whether polymorphisms in CYP1A1, CYP1B1, GSTM1 and GSTT1 act as confounders or as modifiers of those relationships. We performed a secondary analysis of 499 histologically confirmed BC cases and the same number of age-matched population controls. We measured urinary concentrations of 18 metals with mass spectrometry. We determined the genetic variants of interest by allelic discrimination and multiplex PCR. After adjusting for covariates, we found BC negatively associated with arsenic, barium, cobalt, copper, magnesium, molybdenum and vanadium concentrations and positively with those of caesium, manganese, tin and thallium. Most associations remained after stratifying by the genetic variants. We identified that polymorphisms in CYP1B1, CYP1A1 and GSTM1 genes interacted with some metals on BC: interaction p-values CYP1B1 G119T × antimony= 0.036, CYP1B1 G119T × cobalt <0.001, CYP1B1 G119T × tin= 0.032, CYP1A1 A4889G × aluminium= 0.018, CYP1A1 A4889G × arsenic= 0.031, CYP1A1 A4889G × nickel= 0.036, CYP1A1 A4889G × vanadium= 0.031 and GSTM1 deletion × barium= 0.035. Exposure to various individual metals, along with genetic characteristics may contribute to BC development. Further studies are warranted to confirm our results.
Asunto(s)
Neoplasias de la Mama , Exposición a Riesgos Ambientales , Metales , Femenino , Humanos , Arsénico , Bario , Neoplasias de la Mama/genética , Neoplasias de la Mama/epidemiología , Estudios de Casos y Controles , Cobalto , Citocromo P-450 CYP1A1/genética , Exposición a Riesgos Ambientales/efectos adversos , Predisposición Genética a la Enfermedad , Genotipo , Glutatión Transferasa/genética , Metales/efectos adversos , México , Estaño , VanadioRESUMEN
Anthropogenic activities in coastal regions cause risks to the environmental and human health. Due to the carcinogenic and mutagenic potential, polycyclic aromatic hydrocarbons (PAH) are considered priority for monitoring. Most of the Brazilian production of Crassostrea gigas oysters are placed in the Bays of Santa Catarina Island. The aim of this study was to evaluate molecular responses (phase I and II of biotransformation and antioxidant defense) of C. gigas from six oyster farming areas potentially contaminated by sanitary sewage in Florianópolis Metropolitan (SC, Brazil): Santo Antônio de Lisboa, Sambaqui, Serraria, Caieira, Tapera, Imaruim. We evaluated the transcript levels of CYP1A1-like, CYP2-like, CYP2AU2-like, CYP356A1, GSTA1A-like, GSTO.4A-like, SULT-like, SOD-like and CAT-like by qRT-PCR. Only oysters from Caieira showed levels of thermotolerant coliforms allowed by the law. Chemicals analyses in soft tissues of oysters showed low to average levels of PAH in all monitored areas. Enhanced transcript levels of phase I (CYP1A1-like, CYP3564A1-like, CYP2-like and CYP2AU2-like) were observed in oysters from Serraria and Imaruí, suggesting higher biotransformation activity in these farming areas. Regarding phase II of biotransformation, GSTO.4A-like was up-regulated in oysters from Imaruí compared to Caieira and Santo Antônio de Lisboa. An upregulation of SOD-like and CAT-like were observed in oysters from Imaruí and Serraria, suggesting that oysters from these sites are facing higher prooxidant conditions compared to other areas. By integrating the biological and chemical data it is suggested that human-derived contaminants are affecting the oyster metabolism in some farming areas.
Asunto(s)
Crassostrea , Hidrocarburos Policíclicos Aromáticos , Contaminantes Químicos del Agua , Animales , Efectos Antropogénicos , Antioxidantes/metabolismo , Acuicultura , Bahías , Brasil , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , Branquias/metabolismo , Hidrocarburos Policíclicos Aromáticos/análisis , Aguas del Alcantarillado/química , Superóxido Dismutasa/metabolismo , Contaminantes Químicos del Agua/análisisRESUMEN
BACKGROUND: Cancer is currently a major public health problem worldwide, with a marked increase of about 70% in the number of expected diagnosed cases over the next two decades. The amount of tobacco and alcohol consumed is calculated based on the subjective information provided by the user. Tobacco exposure can be assessed using the Fagerström Test for Cigarette Dependence (FTCD) and alcohol consumption by the Alcohol Use Disorder Identification Test (AUDIT). MATERIALS AND METHODS: Forty-eight subjects answered the Fagerström, and AUDIT tests and we studied them as likely screening tools for oral cancer and their correlation with the expression of CYP1A1, GSTM1, GSTP1, and GSTT1 genes by the RT-qPCR method. RESULTS: There were significant differences in the AUDIT score and CYP1A1 expression between cancer and control groups. Participants in advanced stages, whether due to tumor size or regional metastasis, showed significant differences in the duration of tobacco use, FTCD, AUDIT score, and CYP1A1 expression when compared to patients in early stages. Among subjects without cancer, we found a significant correlation between participant age and GSTP1 expression. Furthermore, the expression of GSTP1 was significantly correlated with the number of cigarettes smoked per day, duration of tobacco use, and FTCD. CONCLUSIONS: Questionnaires designed to evaluate the degree of tobacco and alcohol exposure and dependence combined with gene expression tests can be useful to assess the risk of developing oral cancer. Furthermore, raising the awareness of individuals regarding their degree of dependence and encouraging them to participate in cessation programs are important educational measures for the prevention of tobacco-related malignancies.
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Alcoholismo , Neoplasias de la Boca , Consumo de Bebidas Alcohólicas , Estudios de Casos y Controles , Citocromo P-450 CYP1A1/genética , Detección Precoz del Cáncer , Expresión Génica , Predisposición Genética a la Enfermedad , Genotipo , Gutatión-S-Transferasa pi/genética , Glutatión Transferasa/genética , Humanos , Neoplasias de la Boca/diagnóstico , Neoplasias de la Boca/genética , Polimorfismo Genético , NicotianaRESUMEN
The toxicological manifestation of many pollutants relies upon their binding to the aryl hydrocarbon receptor (AHR), and it follows a cascade of reactions culminating in an elevated expression of cytochrome P450 (CYP) 1 enzymes. CYP1A1 and CYP1B1 are associated with enhanced carcinogenesis when chronically exposed to certain polyaromatic hydrocarbons, and their inhibition may lead to chemoprevention. We evaluated dibenzyl trisulfide (DTS), expressed in the ethnomedical plant, Petiveria alliacea, for such potential chemoprevention. Using recombinant human CYP1A1 and CYP1B1 bactosomes on a fluorogenic assay, we first demonstrated that DTS moderately inhibited both enzymes with half maximal inhibitory concentration (IC50) values of 1.3 ± 0.3 and 1.7 ± 0.3 µM, respectively. Against CYP1A1, DTS was a reversible, competitive inhibitor with an apparent inhibitory constant (Ki) of 4.55 ± 0.37 µM. In silico molecular modeling showed that DTS binds with an affinity of -39.8 kJ·mol-1, situated inside the binding pocket, approximately 4.3 Å away from the heme group, exhibiting interactions with phenylalanine residue 123 (Phe-123), Phe-224, and Phe-258. Lastly, zebrafish (Danio rerio) embryos were exposed to 0.08-0.8 µM DTS from 24 to 96 h post fertilization (hpf) with the in vivo ethoxyresorufin-O-deethylase (EROD) assay, and, at 96 hpf, DTS significantly suppressed EROD CYP1A activity in a dose-dependent manner, with up to 60% suppression in the highest 0.8 µM exposure group. DTS had no impact on gene transcription levels for cyp1a and aryl hydrocarbon receptor 2 (ahr2). In co-exposure experiments, DTS suppressed CYP1A activity induced by both B[a]P and PCB-126, although these reductions were not significant. Taken together, these results demonstrate that DTS is a direct, reversible, competitive inhibitor of the carcinogen-activating CYP1A enzyme, binding in the active site pocket close to the heme site, and shows potential in chemoprevention.
Asunto(s)
Compuestos de Bencilo/farmacología , Citocromo P-450 CYP1A1/antagonistas & inhibidores , Citocromo P-450 CYP1B1/antagonistas & inhibidores , Inhibidores Enzimáticos del Citocromo P-450/farmacología , Receptores de Hidrocarburo de Aril/metabolismo , Sulfuros/farmacología , Proteínas de Pez Cebra/metabolismo , Activación Metabólica , Animales , Benzo(a)pireno/metabolismo , Benzo(a)pireno/toxicidad , Compuestos de Bencilo/metabolismo , Sitios de Unión , Unión Competitiva , Dominio Catalítico , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , Citocromo P-450 CYP1B1/genética , Citocromo P-450 CYP1B1/metabolismo , Inhibidores Enzimáticos del Citocromo P-450/metabolismo , Regulación de la Expresión Génica , Humanos , Bifenilos Policlorados/metabolismo , Bifenilos Policlorados/toxicidad , Unión Proteica , Receptores de Hidrocarburo de Aril/genética , Sulfuros/metabolismo , Pez Cebra/embriología , Pez Cebra/metabolismo , Proteínas de Pez Cebra/genéticaRESUMEN
BACKGROUND: Indoleamine 2, 3-dioxygenase-1 (IDO1) is a promising target for immunotherapy in bladder cancer (BC). IDO1 breaks-down tryptophan to generate kynurenine derivatives, which may activate the aryl hydrocarbon receptor (AHR). AHR is an important target for carcinogens, but its association with BC progression was unknown. Two IDO1 inhibitors used in clinical trials are 1-methyl-D-tryptophan (MT) and INCB240360. Because MT is an aromatic hydrocarbon, it may be a ligand for AHR. We hypothesized that AHR could be associated with BC progression and that MT could activate AHR in BC. METHODS: BC patients (n = 165) were selected from the Gene Expression Omnibus database. A cut-off point for relative expression of AHR and cytochrome 450 enzymes (CYP1A1, CYP1A2, and CYP1B1; markers of AHR activation) was determined to compare with the grade, stage, and tumor progression. For in vitro experiments, RT4 (grade 1) and T24 (grade 3) BC cells were incubated with MT and INCB240360 to evaluate the expression of AHR and CYP1A1. RESULTS: AHR activation was associated with grade, stage, and progression of BC. T24 cells express more CYP1A1 than RT4 cells. Although IDO1 expression and kynurenine production are elevated in T24 cells concomitantly to CYP1A1 expression, IDO1 inhibitors were not able to decrease CYP1A1 expression, in contrast, MT significantly increased it in both cell lines. CONCLUSION: In conclusion, it is rational to inhibit IDO1 in BC, among other factors because it contributes to AHR activation. However, MT needs to be carefully evaluated for BC because it is an AHR pathway agonist independently of its effects on IDO1.
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Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/genética , Indolamina-Pirrol 2,3,-Dioxigenasa/genética , Receptores de Hidrocarburo de Aril/genética , Triptófano/análogos & derivados , Neoplasias de la Vejiga Urinaria/tratamiento farmacológico , Anciano , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/antagonistas & inhibidores , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/sangre , Línea Celular Tumoral , Citocromo P-450 CYP1A1/sangre , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A2/sangre , Citocromo P-450 CYP1A2/genética , Citocromo P-450 CYP1B1/sangre , Citocromo P-450 CYP1B1/genética , Progresión de la Enfermedad , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Inmunoterapia , Indolamina-Pirrol 2,3,-Dioxigenasa/antagonistas & inhibidores , Indolamina-Pirrol 2,3,-Dioxigenasa/sangre , Quinurenina/metabolismo , Masculino , Persona de Mediana Edad , Receptores de Hidrocarburo de Aril/antagonistas & inhibidores , Receptores de Hidrocarburo de Aril/sangre , Transducción de Señal/efectos de los fármacos , Triptófano/farmacología , Neoplasias de la Vejiga Urinaria/sangre , Neoplasias de la Vejiga Urinaria/genética , Neoplasias de la Vejiga Urinaria/patologíaRESUMEN
The biotransformation ability of the organism is the result of organ-specific responses. This paper presents a molecular and biochemical approach to elucidate the biotransformation mechanisms in different organs of Prochilodus lineatus induced at 6, 24, and 96â¯h after a benzo[a]pyrene (B[a]P) injection. The induction in cyp1a transcription showed an organ-specific intensity at every tested time time. The EROD (ethoxyresorufin-O-deethylase) activity increased rapidly (6â¯h) in the liver and the kidney; the gills and the brain showed an increase at 24â¯h; and the gills demonstrated the highest activity among all the organs tested. There was no increase in glutathione S-transferase (GST) activity or lipoperoxidation. The decreased hepatic glutathione content (GSH) may be due to its role as an antioxidant. B[a]P was detected in the bile, confirming the xenobiotic efflux from the metabolizing organs. The gills, liver, brain, and kidney of P. lineatus presented an integrated mechanism to deal with the xenobiotic biotransformation.
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Benzo(a)pireno/farmacología , Characiformes/genética , Characiformes/metabolismo , Citocromo P-450 CYP1A1/genética , Animales , Bilis/química , Biotransformación , Encéfalo/metabolismo , Proteínas de Peces/genética , Expresión Génica/efectos de los fármacos , Branquias/metabolismo , Glutatión/metabolismo , Glutatión Transferasa/metabolismo , Riñón/metabolismo , Hígado/metabolismoRESUMEN
The fungicide carbendazim (CBM) has been applied all around the world but its potential adverse effects other than its recognized activity as endocrine disruptor in non target organisms have been scarcely studied. The aims of this work were (1) to use a battery of biomarkers that can reflect potential negative effects such as oxidative stress, genotoxicity, neurotoxicity or altered immune response; and (2) to examine biomarkers of detoxification by analyzing the gene expression of cytochrome P4501A1 (CYP1A1) and the multi-xenobiotic resistance protein P-glycoprotein (P-gp) in the freshwater fish Jenynsia multidentata exposed to environmentally relevant concentrations of CBM during 24 h. Fish exposed to 5 µg/L showed inhibition of GST activity and an increase of TBARs contents in gills, the organ of direct contact with waterborne contaminants. Genotoxicity - measured in peripheral blood-was evidenced by the increases of micronuclei frequency when fish were exposed to 5, 10 and 100 µg/L CBM and of nuclear abnormalities (NA) frequency at 0.05, 0.5, 5, 10 and 100 µg/L CBM. The expression inhibition of interleukin (IL-1ß) and tumor necrosis factor a (TNF-α) at 10, and 5 and 10 µg/L CBM, respectively, indicated an altered immune response. The expression of CYP1A1 was down regulated in liver at 10 µg/L and of P-gp at 5 µg/L CBM, indicating a possible slow on CBM metabolization. On the other hand, in gills CYP1A1 decreased at 5 and 10 µg/L while P-gp was induced at 5 and 100 µg/L CBM. Overall, most of these significant effects were detected below 10 µg/L CBM, in a range of realistic concentrations in aquatic ecosystems worldwide.
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Bencimidazoles/toxicidad , Carbamatos/toxicidad , Ciprinodontiformes/metabolismo , Citocinas/metabolismo , Micronúcleos con Defecto Cromosómico/inducido químicamente , Estrés Oxidativo/efectos de los fármacos , Contaminantes Químicos del Agua/toxicidad , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/metabolismo , Animales , Antioxidantes/metabolismo , Biomarcadores/metabolismo , Ciprinodontiformes/genética , Ciprinodontiformes/inmunología , Citocromo P-450 CYP1A1/genética , Ecosistema , Agua Dulce/química , Branquias/efectos de los fármacos , Branquias/metabolismo , Hígado/efectos de los fármacos , Hígado/metabolismo , Estrés Oxidativo/genética , Estrés Oxidativo/inmunologíaRESUMEN
Lubricant oils are among oil-based products that are not fully consumed during its use, thereby producing non-biodegradable residues which can cause contamination of natural systems. This study evaluated the toxicity of new and used lubricating oil (0.01 and 0.1 mL L-1) in adult Nile tilapia (Oreochromis niloticus), by assessing the effects on oxidative stress, biotransformation enzymes (liver and gills), and histopathological alterations on hepatic and pancreatic tissues after 3 and 7 days of exposure. Results showed that 3-days exposure to 0.1 mL L-1 of used and new lubricating oil increased the activity of superoxide dismutase (SOD) and malondialdehyde (MDA) levels in liver of O. niloticus, respectively. In gills, catalase (CAT) was decreased in fish exposed to 0.1 mL L-1 of non-used oil after 3 days, but pronounced increases in CAT was detected after 7 days-exposure to both new and used oil. Shorter exposure to both concentrations of new and used oil also raised glutathione-S-transferase activity (GST) in gills. Ethoxyresorufin-O-deethylase (EROD) was induced in liver of fish exposed to 0.1 mL L-1of used oil after 3 and 7 days, however a reduced response of this enzyme was detected in gills of animals from both oil treatments. In vitro analysis showed that hepatic EROD was inhibited by lubricating oil exposures, with more pronounced responses in treatments containing used oil. Hepatic lesions, such as cytoplasmic vacuolization, nuclei abnormally, changes in hepatocytes shape, steatosis, cholestasis, eosinophilic inclusions and necrosis were mainly increased by 7 days exposure to used lubricating oil at higher concentration.
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Cíclidos/fisiología , Gasolina/toxicidad , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/patología , Lubricantes/toxicidad , Estrés Oxidativo/efectos de los fármacos , Animales , Automóviles , Biotransformación/efectos de los fármacos , Catalasa/genética , Catalasa/metabolismo , Cíclidos/genética , Cíclidos/metabolismo , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , Branquias/efectos de los fármacos , Branquias/patología , Glutatión Transferasa/genética , Glutatión Transferasa/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Masculino , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismoRESUMEN
Coal plants represent one of the main sources of environmental pollution due to the combustion process of this mineral and the consequent release of gases and particles which, in significant quantities, can lead to a potential risk to health and the environment. The susceptibility of individuals to the genotoxic effects of coal mining can be modulated by genetic variations in the xenobiotic detoxification and DNA repair processes. The aim of this study was to evaluate if xenobiotic metabolism polymorphism, base excision repair polymorphisms and non-homologous end joining repair polymorphism, could modify individual susceptibility to genomic instability and epigenetic alterations induced in workers by occupational exposure to coal. In this study, polymerase chain reaction was used to examine the polymorphic sites. The sample population comprising 70 coal mine workers and 71 workers non-exposed to coal. Our results demonstrated the effect of individual genotypes on different biomarkers evaluated. Significant decrease in % of global DNA methylation were observed in CYP1A1 Val/- exposed individuals compared to CYP1A1 Ile/Ile individuals. Coal workers who carried the XRCC4 Ile/Ile genotype showed decrease NBUD frequencies, while the XRCC4 Thr/- genotype was associated with decrease in Buccal micronucleus cells for the group not exposed. No influence of GSTM1 null, GSTT1 null, GSTP1 Ile105Val, hOGG1 Ser326Cys, XRCC1 Arg194Trp polymorphisms was observed. Thus, the current study reinforces the importance of considering the effect of metabolizing and repair variant genotypes on the individual susceptibility to incorporate DNA damage, as these processes act in a coordinated manner to determine the final response to coal exposure.
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Minas de Carbón , Carbón Mineral/toxicidad , Daño del ADN , Metilación de ADN , Exposición Profesional , Polimorfismo Genético , Homeostasis del Telómero , Adolescente , Adulto , Anciano , Citocromo P-450 CYP1A1/genética , Reparación del ADN , Proteínas de Unión al ADN/genética , Femenino , Genotipo , Gutatión-S-Transferasa pi/genética , Glutatión Transferasa/genética , Humanos , Masculino , Persona de Mediana Edad , Proteína 1 de Reparación por Escisión del Grupo de Complementación Cruzada de las Lesiones por Rayos X/genética , Xenobióticos/metabolismo , Adulto JovenRESUMEN
Polycyclic aromatic hydrocarbons (PAHs) are among the main contaminants in aquatic environments. PAHs can affect organisms due to their carcinogenic, mutagenic and/or teratogenic characteristics. Depending on the PAHs, concentration, and period of exposure, biological damage can occur leading to histopathologic alterations. This study aimed to evaluate the molecular, biochemical and histological responses of the oyster Crassostrea gasar exposed to pyrene (0.25 and 0.5⯵M) and fluorene (0.6 and 1.2⯵M), after exposure for 24 and 96â¯h. Concentrations of both PAHs were quantified in the water and in oyster tissues. Transcript levels of phase I (CYP3475C1, CYP2-like, CYP2AU1 and CYP356A) and phase II (GSTO-like, MGST-like and SULT-like) biotransformation-related genes and the activities of ethoxyresorufin-O-deethylase (EROD), total and microsomal glutathione S-transferase (GST and MGST) were evaluated in the gills. Also, histological changes and localization of mRNA transcripts CYP2AU1 in gills, mantle, and digestive diverticula were evaluated. Both PAHs accumulated in oyster tissues. Pyrene half-life in water was significantly lower than fluorene. Transcript levels of all genes were higher in oysters exposed to of pyrene 0.5⯵M (24â¯h). Only CYP2AU1 gene was up-regulated by fluorene exposure. EROD and MGST activities were higher in oysters exposed to pyrene. Tubular atrophy in the digestive diverticula and an increased number of mucous cells in the mantle were observed in oysters exposed to pyrene. CYP2AU1 transcripts were observed in different tissues of pyrene-exposed oysters. A significant correlation was observed between tubular atrophy and the CYP2AU1 hybridization signal in oysters exposed to pyrene, suggesting the sensibility of the species to this PAH. These results suggest an important role of biotransformation-related genes and enzymes and tissue alterations associated to pyrene metabolism but not fluorene. In addition, it reinforces the role of CYP2AU1 gene in the biotransformation process of PAHs in the gills of C. gasar.
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Crassostrea/citología , Crassostrea/genética , Fluorenos/toxicidad , Pirenos/toxicidad , Animales , Biotransformación/efectos de los fármacos , Crassostrea/efectos de los fármacos , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , Sistema Digestivo/efectos de los fármacos , Fluorescencia , Regulación de la Expresión Génica/efectos de los fármacos , Branquias/efectos de los fármacos , Branquias/enzimología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Contaminantes Químicos del Agua/toxicidadRESUMEN
Parkin (PRKN) is a ubiquitin E3 ligase that catalyzes the ubiquitination of several proteins. Mutations in the human Parkin gene, PRKN, leads to degeneration of dopaminergic (DA) neurons, resulting in autosomal recessive early-onset parkinsonism and the loss of PRKN function is linked to sporadic Parkinson's disease (PD). Additionally, several in vitro studies have shown that overexpression of exogenous PRKN protects against the neurotoxic effects induced by a wide range of cellular stressors, emphasizing the need to study the mechanism(s) governing PRKN expression and induction. Here, Prkn was identified as a novel target gene of the aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor and member of the bHLH/PAS (basic helix-loop-helix/Per-Arnt-Sim) superfamily. AhR binds and transactivates the Prkn gene promoter. We also demonstrated that AhR is expressed in DA neurons and that its activation upregulates Prkn mRNA and protein levels in the mouse ventral midbrain. Additionally, the AhR-dependent increase in PRKN levels is associated with a decrease in the protein levels of its target substrate, α-synuclein, in an AhR-dependent manner, because this effect is not observed in Ahr-null mice. These results suggest that treatments designed to induce PRKN expression through the use of nontoxic AhR agonist ligands may be novel strategies to prevent and delay PD.
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Ubiquitina-Proteína Ligasas/metabolismo , alfa-Sinucleína/metabolismo , Actinas/metabolismo , Animales , Encéfalo/metabolismo , Línea Celular , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , Regulación de la Expresión Génica/fisiología , Humanos , Hígado/metabolismo , Ratones , Ratones Noqueados , Neuronas/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptores de Hidrocarburo de Aril/metabolismo , Ubiquitina-Proteína Ligasas/genética , alfa-Sinucleína/genéticaRESUMEN
BACKGROUND: Pathogens stimulate immune functions of macrophages. Macrophages are a key sentinel cell regulating the response to pathogenic ligands and orchestrating the direction of the immune response. Our study aimed at investigating the early transcriptomic changes of bovine macrophages (Bomacs) in response to stimulation with CpG DNA or polyI:C, representing bacterial and viral ligands respectively, and performed transcriptomics by RNA sequencing (RNASeq). KEGG, GO and IPA analytical tools were used to reconstruct pathways, networks and to map out molecular and cellular functions of differentially expressed genes (DE) in stimulated cells. RESULTS: A one-way ANOVA analysis of RNASeq data revealed significant differences between the CpG DNA and polyI:C-stimulated Bomac. Of the 13,740 genes mapped to the bovine genome, 2245 had p-value ≤0.05, deemed as DE. At 6 h post stimulation of Bomac, poly(I:C) induced a very different transcriptomic profile from that induced by CpG DNA. Whereas, 347 genes were upregulated and 210 downregulated in response to CpG DNA, poly(I:C) upregulated 761 genes and downregulated 414 genes. The topmost DE genes in poly(I:C)-stimulated cells had thousand-fold changes with highly significant p-values, whereas in CpG DNA stimulated cells had 2-5-fold changes with less stringent p-values. The highest DE genes in both stimulations belonged to the TNF superfamily, TNFSF18 (CpG) and TNFSF10 (poly(I:C)) and in both cases the lowest downregulated gene was CYP1A1. CpG DNA highly induced canonical pathways that are unrelated to immune response in Bomac. CpG DNA influenced expression of genes involved in molecular and cellular functions in free radical scavenging. By contrast, poly(I:C) highly induced exclusively canonical pathways directly related to antiviral immune functions mediated by interferon signalling genes. The transcriptomic profile after poly(I:C)-stimulation was consistent with induction of TLR3 signalling. CONCLUSION: CpG DNA and poly(I:C) induce different early transcriptional landscapes in Bomac, but each is suited to a specific function of macrophages during interaction with pathogens. Poly(I:C) influenced antiviral response genes, whereas CpG DNA influenced genes important for phagocytic processes. Poly(I:C) was more potent in setting the inflammatory landscape desirable for an efficient immune response against virus infection.
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Secuenciación de Nucleótidos de Alto Rendimiento , Macrófagos/metabolismo , Moléculas de Patrón Molecular Asociado a Patógenos , Transcriptoma/genética , Animales , Bovinos , Línea Celular , Islas de CpG/genética , Citocromo P-450 CYP1A1/genética , Perfilación de la Expresión Génica , Genoma/genética , Ligandos , Macrófagos/microbiología , Macrófagos/virología , Poli I-C/genética , Factores de Necrosis Tumoral/genéticaRESUMEN
The goal of this work was to design specific cyp1a primers for the fish Prochilodus lineatus to study the expression of this gene and its relation to the activity of biotransformation phase I enzyme (ethoxyresorufin O-deethylase - EROD) and genotoxic damage after 6 and 24â¯h of benzo(a)pyrene (B(a)P) intraperitoneal injection. In comparison to fish injected only with canola oil (vehicle), the expression of cyp1a and EROD activity both in the liver and gills were significantly higher after 6 and 24â¯h of B(a)P injection. A significant increase in DNA damage was detected in liver and blood cells after 6â¯h of B(a)P injection and in the gill cells after both times, probably caused by intermediate metabolites of B(a)P. Thus, the expression of cyp1a and its relationship with the corresponding enzyme activity is a potential biomarker for evaluation P. lineatus exposure to organic compounds.
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Benzo(a)pireno/toxicidad , Characiformes , Citocromo P-450 CYP1A1/genética , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Contaminantes Químicos del Agua/toxicidad , Animales , Biomarcadores/metabolismo , Characiformes/genética , Characiformes/metabolismo , Citocromo P-450 CYP1A1/metabolismo , Daño del ADN , Eritrocitos/efectos de los fármacos , Eritrocitos/metabolismo , Branquias/efectos de los fármacos , Branquias/metabolismo , Glutatión/metabolismo , Glutatión Transferasa/metabolismo , Hígado/efectos de los fármacos , Hígado/metabolismo , Malondialdehído/metabolismoRESUMEN
PURPOSE: During recent decades, several reports have suggested a decrease in semen quality and DNA damage due in part to environmental toxicants and industrial chemicals. Among these xenobiotics, polycyclic aromatic hydrocarbons (PAHs) are of particular concern because of their remarkable mutagenic and carcinogenic properties and because several experimental and epidemiological studies have reported adverse effects of PAHs on male reproductive health and DNA structure. The aim of the study was to evaluate the association between 1-hydroxypyrene (1-OHP) urinary levels and sperm quality, DNA damage and the frequency of CYP1A1, GSTT1, and GSTM1 polymorphisms. METHODS: Semen, urine and blood samples were taken for sperm-quality assessment, 1-OHP urinary level measurement, DNA damage evaluation and polymorphism frequency analysis of three genes implicated in PAH metabolism in a total of 70 Mexican subjects exposed and nonexposed to PAHs. RESULTS: A significant decrease in sperm quality and increased DNA damage were registered in occupationally exposed volunteers. Polymorphisms modified the 1-OHP urinary levels; however, no associations were found between them. Inverse associations were registered between the sperm concentration/mL and 1-OHP levels and between tail lengths and the GSMT1 null genotype. CONCLUSIONS: Our data showed an inverse association between 1-OHP urinary levels and both sperm quality and the DNA integrity. Additionally, the heterozygote variants of CYP1A1-m1 and CYP1A1-m2 significantly increased the urinary excretion of 1-OHP, and the GSTM1 null variant was inversely associated with the comet parameters evaluated.