RESUMEN
Microbiota alterations are observed in pathological conditions, and their regulation is a subject of great interest. Gut microbes are affected by diet, and plant polyphenols may have positive effect on gut microbiota; however, plant-derived extracts may have toxic effects. Guarana (Paullinia cupana Mart.) is a nontraditional medicinal plant applied worldwide. Guarana yields an alkaloid and polyphenol-rich seed with antimicrobial, antioxidant, and anti-inflammatory properties, where caffeine is the major compound. We evaluated the effects of guarana seed powder (GSP) and purified caffeine on gut microbial composition and redox and inflammatory parameters in Wistar rats after 21 days of treatment. Fecal microbiota was analyzed utilizing 16S rDNA sequencing. Antioxidant enzymes activities from liver, kidney, and colon, as well as oxidative damage markers, were evaluated. Total nonenzymatic antioxidant potential was also evaluated. Microbiota was altered by both treatments, GSP and caffeine, without loss of diversity. In the liver, the kidney, and the colon, we observed a decrease in the antioxidant enzymes activities in the GSP group with no increase in the expression of oxidative damage markers, although some enzymes were also regulated by caffeine. Taken together, these results suggested that GSP ameliorates redox parameters but negatively affected gut microbiota, partially via caffeine.
Asunto(s)
Cafeína/farmacología , Microbioma Gastrointestinal/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Teobromina/farmacología , Teofilina/farmacología , Animales , Antioxidantes/farmacología , Cafeína/química , Disbiosis/inducido químicamente , Disbiosis/microbiología , Disbiosis/patología , Masculino , Oxidación-Reducción/efectos de los fármacos , Paullinia/química , Extractos Vegetales/farmacología , Plantas Medicinales/química , Ratas , Ratas Wistar , Semillas , Teobromina/química , Teofilina/químicaRESUMEN
BACKGROUND/AIMS: Heat shock protein 70 (HSP70) has been recently described with extracellular actions, where it is actively released in inflammatory conditions. Acting as DAMPs (damage associated molecular pattern), extracellular HSP70 (eHSP70) interacts with membrane receptors and activates inflammatory pathways. At this context, the receptor for advanced glycation endproducts (RAGE) emerges as a possible candidate for interaction with eHSP70. RAGE is a pattern-recognition receptor and its expression is increased in several diseases related to a chronic pro-inflammatory state. One of the main consequences of RAGE ligand-binding is the ERK1/2 (extracellular signal-regulated kinases)-dependent activation of NF-kB (nuclear factor kappa B), which leads to expression of TNF-α (tumor necrosis factor alpha) and other cytokines. The purpose of this work is to elucidate if eHSP70 is able to evoke RAGE-dependent signaling using A549 human lung cancer cells, which constitutively express RAGE. METHODS: Immunoprecipitation and protein proximity assay were utilized to demonstrate the linkage between RAGE and eHSP70. To investigate RAGE relevance on cell response to eHSP70, siRNA was used to knockdown the receptor expression. Signaling pathways activation were evaluated by western blotting, gene reporter luciferase and real time quantitative PCR. RESULTS: Protein eHSP70 shown to be interacting physically with the receptor RAGE in our cell model. Treatment with eHSP70 caused ERK1/2 activation and NF-κB transactivation impaired by RAGE knockdown. Moreover, the stimulation of pro-inflammatory cytokines expression by eHSP70 was inhibited in RAGE-silenced cells. Finally, conditioned medium of eHSP70-treated A549 cells caused differential effects in monocytes cytokine expression when A549 RAGE expression is inhibited. CONCLUSIONS: Our results evidence eHSP70 as a novel RAGE agonist capable of influence the cross-talk between cancer and immune system cells.
Asunto(s)
Proteínas HSP70 de Choque Térmico/metabolismo , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , FN-kappa B/metabolismo , Receptor para Productos Finales de Glicación Avanzada/metabolismo , Células A549 , Citocinas/genética , Citocinas/metabolismo , Proteínas HSP70 de Choque Térmico/genética , Proteínas HSP70 de Choque Térmico/farmacología , Humanos , Immunoblotting , Inmunoprecipitación , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , FN-kappa B/genética , Fosforilación , Interferencia de ARN , ARN Interferente Pequeño/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptor para Productos Finales de Glicación Avanzada/antagonistas & inhibidores , Receptor para Productos Finales de Glicación Avanzada/genética , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/farmacología , Transducción de Señal/efectos de los fármacos , Activación Transcripcional , Células U937RESUMEN
Most scientific studies are too long to be conducted in a single day or even in a few days. Thus, there is a need to store samples for subsequent investigations. There is sparse information about specific sample storage protocols that minimize analytical error and variability in evaluations of redox parameters. Therefore, the effects of storage temperature and freezing time on enzymatic activities, protein oxidative damage, and CAT (catalase) and SOD1 (superoxide dismutase) immunocontent of blood, liver, and brain from rats were determined for two different sample forms (frozen homogenized tissue or frozen intact tissue). Superoxide dismutase activity was drastically decreased in blood and liver with an increase in freezing time, but not in brain. Catalase activity showed a decrease only in intact liver at -20 and -80°C. In contrast, in blood it showed an increase in intact tissue at -20 and -80°C. Reduced thiol groups generally decreased with freezing time, but showed an increase in intact blood at -20 and -80°C, probably because of color interference. Carbonyl groups in homogenized liver and brain, and in intact blood (except at 80°C) drastically increased with freezing time. Freezing time did not modulate the immunocontent of CAT and SOD1 levels in any tissue. In conclusion, our results indicate that storage at -20°C affects redox parameters more than storage at -80°C. Storage for a long time may compromise the samples, leading to changing parameters due to oxidative stress. Thus, we suggest processing the samples as soon as possible. However, if this is not possible, then material can be aliquoted into different tubes to prevent the effect of refreezing of samples.
Asunto(s)
Encéfalo/enzimología , Congelación , Hígado/enzimología , Manejo de Especímenes/normas , Animales , Encéfalo/inmunología , Catalasa/sangre , Catalasa/inmunología , Catalasa/metabolismo , Calefacción , Hígado/inmunología , Oxidación-Reducción , Ratas , Superóxido Dismutasa/sangre , Superóxido Dismutasa/inmunología , Superóxido Dismutasa/metabolismoRESUMEN
Early life stress (ELS) has been associated with biological and psychosocial alterations due to developmental reprogramming. Here, we investigated whether childhood maltreatment is associated with an imbalance between the production of oxidative markers and antioxidant defenses. Thirty adolescents with no psychiatric disorder but reporting childhood maltreatment and twenty-seven adolescents with no psychiatric disorder and no history of ELS were recruited for the study. Childhood maltreatment was investigated by the Childhood Trauma Questionnaire (CTQ). Redox state was estimated by plasma levels of protein carbonylation, total thiol content (SH), superoxide dismutase (SOD), glutathione peroxidase (GPx), as well as total reactive antioxidant potential (TRAP). Childhood maltreatment was associated with oxidative stress as shown by increased protein carbonylation. Interestingly, adolescents exposed to maltreatment also displayed higher SOD levels, TRAP kinetics and reduced GPx levels when compared with adolescents who had not undergone childhood maltreatment. No significant differences were observed for SH levels. Taken together, we provide novel evidence indicating that childhood maltreatment is associated with increased oxidative stress markers in otherwise healthy adolescents.
Asunto(s)
Antioxidantes/metabolismo , Maltrato a los Niños , Estrés Oxidativo , Estrés Psicológico/metabolismo , Adolescente , Femenino , Glutatión Peroxidasa/sangre , Humanos , Masculino , Oxidación-Reducción , Carbonilación Proteica , Especies Reactivas de Oxígeno/sangre , Superóxido Dismutasa/sangreRESUMEN
RATIONALE: Preclinical studies have shown that cocaine exposure and withdrawal are associated with cellular oxidative stress damage. However, the impact of crack-cocaine dependence on oxidative stress biomarkers remains unclear. Here, we assessed peripheral oxidative stress and antioxidant defences during two periods of crack-cocaine detoxification treatment and associated these changes with psychological morbidity. METHODS: Thirty female inpatients were recruited, and plasma samples were collected at the 4th and 18th days of abstinence; 30 healthy controls were also recruited. Plasma levels of protein carbonyl, protein thiol content, superoxide dismutase (SOD), glutathione peroxidase (GPx), reduced reduced (GSH) and total reactive antioxidant potential (TRAP) were measured by standard methods; the questionnaires Cocaine Selective Severity Assessment, Beck Depressive Inventory and the Addiction Severity Index were applied. RESULTS: We report higher oxidative stress damage after 4 days of detoxification, as shown by increased total thiol content and protein carbonylation when compared with control group and after 18 days of detoxification. After 18 days of treatment, we observed a recovery of the oxidative stress damage and increase of the antioxidant defences, as shown by higher levels of SOD, GPx, GSH and TRAP. There was a positive correlation between protein carbonylation and psychological variables; in contrast, there was a negative correlation between TRAP levels and clinical assessments. CONCLUSIONS: Taken together, these results suggest that drug rehabilitation treatment was effective in decreasing oxidative damage represented by the reduction in biological markers, which are closely related to the severity of withdrawal symptoms.
Asunto(s)
Antioxidantes/metabolismo , Trastornos Relacionados con Cocaína/sangre , Trastornos Relacionados con Cocaína/terapia , Cocaína Crack , Estrés Oxidativo/fisiología , Adulto , Trastornos Relacionados con Cocaína/diagnóstico , Cocaína Crack/efectos adversos , Femenino , Estudios de Seguimiento , Glutatión Peroxidasa/sangre , Humanos , Síndrome de Abstinencia a Sustancias/sangre , Síndrome de Abstinencia a Sustancias/diagnóstico , Síndrome de Abstinencia a Sustancias/terapia , Superóxido Dismutasa/sangre , Encuestas y Cuestionarios , Factores de Tiempo , Resultado del Tratamiento , Adulto JovenRESUMEN
Vitamin A is important for both development and maintenance of adult brain homeostasis. However, excessive vitamin A exposure has been linked to cognitive impairments and may induce congenital defects, including neuronal malformations. Recently, we demonstrated that vitamin A supplementation is able to alter behavioral parameters and induce a pro-oxidant state in hippocampus and striatum of adult male rat. Thus, the aim of the present work was to investigate the effects of vitamin A supplementation in pregnant and nursing rats on maternal and offspring striatum and hippocampus. Wistar female rats (7 per group) were orally supplemented with retinyl palmitate (2500, 12,500 and 25,000 IU/kg/day) or saline (control) throughout pregnancy and nursing. Homing test was performed at postnatal days (PND) 5 and 10 for offspring, while open field test (OFT) was carried out at PND19 and 20 for dams and offspring, respectively. Redox parameters were evaluated at PND21 for both. Vitamin A supplementation during pregnancy and nursing increased superoxide dismutase/catalase (SOD/CAT) ratio and oxidative damage in maternal and offspring striatum and hippocampus. Additionally, supplementation induced behavioral alterations. In conclusion, we suggest some caution regarding vitamin A intake during pregnancy and breastfeeding, since oxidative stress can disturb several biological phenomena, including neuronal signaling and neurotransmission, which may induce several behavioral deficits.