RESUMEN
Gastroesophageal reflux disease (GERD), a clinical condition characterized by reflux of gastroduodenal contents in the oesophagus, has proved to demonstrate a strong link between oxidative stress and the development of GERD. Proton pump inhibitors (PPIs) have been universally accepted as first-line therapy for management of GERD. The potential benefits of electrolysed reduced water (ERW), rich in molecular hydrogen, in improving symptoms and systemic oxidative stress associated with GERD was assessed. The study was performed on 84 GERD patients undergoing control treatment (PPI + tap water) or experimental treatment (PPI + ERW) for 3 months. These patients were subjected to the GERD-Health Related Quality of Life Questionnaire as well as derivatives reactive oxigen metabolites (d-ROMs) test, biological antioxidant potential (BAP) test, superoxide anion, nitric oxide and malondialdehyde assays, which were all performed as a proxy for the oxidative/nitrosative stress and the antioxidant potential status. Spearman's correlation coefficient was used to evaluate the correlation between scores and laboratory parameters. Overall results demonstrated that an optimal oxidative balance can be restored and GERD symptoms can be reduced rapidly via the integration of ERW in GERD patients. The relative variation of heartburn and regurgitation score was significantly correlated with laboratory parameters. Thus, in the selected patients, combination treatment with PPI and ERW improves the cellular redox state leading to the improvement of the quality of life as demonstrated by the correlation analysis between laboratory parameters and GERD symptoms.
Asunto(s)
Reflujo Gastroesofágico/sangre , Reflujo Gastroesofágico/terapia , Hidrógeno/uso terapéutico , Agua/farmacología , Adulto , Anciano , Antioxidantes/metabolismo , Humanos , Persona de Mediana Edad , Oxidación-Reducción , Estrés Oxidativo/efectos de los fármacos , Inhibidores de la Bomba de Protones/uso terapéutico , Calidad de Vida , Adulto JovenRESUMEN
The cognitive impairment characterizing the phenotype of older adults has been related to the efficiency of the antioxidant system. This study aimed at investigating the effect of memory training (MT) on memory, global cognitive functioning, and the oxidant and antioxidant capacity of plasma. We recruited 52 healthy subjects aged over 60. Twenty-nine subjects were submitted to 6-months of MT (Experimental Group, EG), and 23 were used as a Control Group (CG). Global cognitive functioning was assessed by the Mini-Mental State Examination (MMSE) and Short- and Long-Term Memory (STM and LTM, respectively) by the Rey Auditory Verbal Learning Test (RAVLT) at baseline (T0) and after 6-months (T1). Meanwhile, Reactive Oxygen Metabolites derivative compounds (d-ROMs), Biological Antioxidant Potential (BAP), and their ratio were evaluated on plasma. Results showed that the MMSE and RAVLT scores improved in EG at T1. At the same time, the d-ROMs levels significantly decreased, while the BAP and BAP/d-ROMs ratio showed an opposite trend. In both groups, the MMSE and LTM scores were negatively associated with d-ROMs levels, and positively correlated with BAP levels and the BAP/d-ROMs ratio. When we considered the Δvalue (Δvariableâ¯=â¯variable post-MT minus variable pre-MT) in EG, the ΔMMSE and ΔLTM scores were negatively associated to Δd-ROMs, and positively to ΔBAP and ΔBAP/dROM. In conclusion, our results suggest that MT improves memory and global cognitive functioning. These processes were significantly associated to increase in resistance against oxidative stress at the plasma level in healthy older adults.
Asunto(s)
Envejecimiento/sangre , Envejecimiento/psicología , Aprendizaje , Estrés Oxidativo , Anciano , Anciano de 80 o más Años , Biomarcadores/sangre , Femenino , Humanos , Aprendizaje/fisiología , Masculino , Persona de Mediana Edad , Pruebas Neuropsicológicas , Estrés Oxidativo/fisiologíaRESUMEN
It has been shown that functional recovery of patients with acute congestive heart failure (ACHF) after treatment with conventional drugs (CD) is mediated by suppression of inflammation in peripheral blood mononuclear cells. Here, we analyzed gene expression profiles of monocytes from symptomatic ACHF patients (NYHA Class III-IV) before and after pharmacological treatment with CD. The treatment was associated with selective down-regulation of "TNFR signaling" and pro-inflammatory mediators CCL5, MIP-1α receptor, CD14, ITGAM, and significant up-regulation of "TNFR signaling" as evidenced by increase in anti-inflammatory factors including NF-kBIA, TNFAIP3 and SHP-1. In monocyte TNF-alpha-stimulated there is a down-regulation of the phosphatase SHP-1 which induces a significant activation of TAK-1/IKK/NF-kB signaling. These findings suggest that the therapeutic impact of CD treatment in symptomatic ACHF includes negative regulation of the NF-kB signaling in monocytes and the improvement of the SHP-1 activity.
Asunto(s)
Insuficiencia Cardíaca/sangre , Monocitos/metabolismo , FN-kappa B/sangre , Proteína Tirosina Fosfatasa no Receptora Tipo 6/sangre , Anciano , Estudios de Casos y Controles , Femenino , Insuficiencia Cardíaca/genética , Humanos , Quinasa I-kappa B/sangre , Linfocitos/metabolismo , Quinasas Quinasa Quinasa PAM/sangre , Masculino , Persona de Mediana Edad , Neutrófilos/metabolismo , Proteína Tirosina Fosfatasa no Receptora Tipo 6/antagonistas & inhibidores , Proteína Tirosina Fosfatasa no Receptora Tipo 6/genética , ARN Interferente Pequeño/genética , Transducción de Señal , Transcriptoma , Factor de Necrosis Tumoral alfa/sangreRESUMEN
It is known that increased levels of reactive oxygen species (ROS) and reactive nitrogen species (RNS) can exert harmful effects, altering the cellular redox state. Electrolyzed Reduced Water (ERW) produced near the cathode during water electrolysis exhibits high pH, high concentration of dissolved hydrogen and an extremely negative redox potential. Several findings indicate that ERW had the ability of a scavenger free radical, which results from hydrogen molecules with a high reducing ability and may participate in the redox regulation of cellular function. We investigated the effect of ERW on H2O2-induced U937 damage by evaluating the modulation of redox cellular state. Western blotting and spectrophotometrical analysis showed that ERW inhibited oxidative stress by restoring the antioxidant capacity of superoxide dismutase, catalase and glutathione peroxidase. Consequently, ERW restores the ability of the glutathione reductase to supply the cell of an important endogenous antioxidant, such as GSH, reversing the inhibitory effect of H2O2 on redox balance of U937 cells. Therefore, this means a reduction of cytotoxicity induced by peroxynitrite via a downregulation of the NF-κB/iNOS pathway and could be used as an antioxidant for preventive and therapeutic application. In conclusion, ERW can protect the cellular redox balance, reducing the risk of several diseases with altered cellular homeostasis such as inflammation.
Asunto(s)
Antioxidantes/farmacología , FN-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Agua/farmacología , Antioxidantes/química , Línea Celular Tumoral , Electrólisis/métodos , Humanos , Oxidación-Reducción , Estrés Oxidativo/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Investigación Biomédica Traslacional/métodos , Agua/químicaRESUMEN
Several studies have shown that xanthones obtained from Garcinia Mangostana (GM) have remarkable biological activities. α-mangostin (α-MG) is the main constituent of the fruit hull of the GM. Several findings have suggested that SIRT-1, a nuclear histone deacetylase, could influence cellular function by the inhibition of NF-kB signaling. ROS can inhibit SIRT-1 activity by initiating oxidative modifications on its cysteine residues, and suppression of SIRT-1 enhances the NF-κB signaling resulting in inflammatory responses. The goals of the present study were to evaluate the quantity of α-MG in the methanolic extract of GM (Vithagroup Spa) and to investigate the activity of this xanthone in U937 cell line and in human monocytes from responsive to inflammatory insult analyzing the possible changes on the activation of SIRT-1 protein via NF-Kb. Cells were treated with the methanolic extract of GM and/or LPS. The chromatographic separation of α-MG was performed by an HPLC analysis. EX 527, a specific SIRT-1 inhibitor, was used to determine if SIRT-1/NfkB signaling pathway might be involved in α-MG action on cells. Our results show that α-MG inhibits p65 acetylation and down-regulates the pro-inflammatory gene products as COX-2, iNOS via SIRT-1 activation. Cells treated with EX 527 showed an up-regulation of NFkB acetylation and an over expression of inducible enzymes and their product of catalysis (NO and PGE2). These results suggest that α-MG may be useful for the development of alternative pharmacological strategies aimed at reducing the inflammatory process. J. Cell. Physiol. 231: 2439-2451, 2016. © 2016 Wiley Periodicals, Inc.
Asunto(s)
Inflamación/patología , Transducción de Señal/efectos de los fármacos , Sirtuina 1/metabolismo , Xantonas/farmacología , Acetilación/efectos de los fármacos , Muerte Celular/efectos de los fármacos , Citocinas/genética , Citocinas/metabolismo , Citoprotección/efectos de los fármacos , Depuradores de Radicales Libres/farmacología , Garcinia/química , Humanos , Lipopolisacáridos , Monocitos/efectos de los fármacos , Monocitos/metabolismo , FN-kappa B/metabolismo , Extractos Vegetales/farmacología , Superóxidos/metabolismo , Células U937 , Xantonas/químicaRESUMEN
Several reports suggest that ELF-EMF exposures interact with biological processes including promotion of cell proliferation. However, the molecular mechanisms by which ELF-EMF controls cell growth are not completely understood. The present study aimed to investigate the effect of ELF-EMF on keratinocytes proliferation and molecular mechanisms involved. Effect of ELF-EMF (50 Hz, 1 mT) on HaCaT cell cycle and cells growth and viability was monitored by FACS analysis and BrdU assay. Gene expression profile by microarray and qRT-PCR validation was performed in HaCaT cells exposed or not to ELF-EMF. mTOR, Akt and MAPKs expressions were evaluated by Western blot analysis. In HaCaT cells, short ELF-EMF exposure modulates distinct patterns of gene expression involved in cell proliferation and in the cell cycle. mTOR activation resulted the main molecular target of ELF-EMF on HaCaT cells. Our data showed the increase of the canonical pathway of mTOR regulation (PI3K/Akt) and activation of ERK signaling pathways. Our results indicate that ELF-EMF selectively modulated the expression of multiple genes related to pivotal biological processes and functions that play a key role in physio-pathological mechanisms such as wound healing.
Asunto(s)
Queratinocitos/metabolismo , Sistema de Señalización de MAP Quinasas/genética , Fosfatidilinositol 3-Quinasas/genética , Proteínas Proto-Oncogénicas c-akt/genética , Transducción de Señal/genética , Serina-Treonina Quinasas TOR/genética , Técnicas de Cultivo de Célula , Proliferación Celular/genética , Campos Electromagnéticos , Humanos , Masculino , Persona de Mediana Edad , Familia de Multigenes/genética , Transcriptoma/genética , Cicatrización de Heridas/genéticaRESUMEN
Polyphenols are the major components of many traditional herbal remedies, which exhibit several beneficial effects including anti-inflammation and antioxidant properties. Src homology region 2 domain-containing phosphatase-1 (SHP-1) is a redox sensitive protein tyrosine phosphatase that negatively influences downstream signalling molecules, such as mitogen-activated protein kinases, thereby inhibiting inflammatory signalling induced by lipopolysaccharide (LPS). Because a role of transforming growth factor ß-activated kinase-1 (TAK1) in the upstream regulation of JNK molecule has been well demonstrated, we conjectured that SHP-1 could mediate the anti-inflammatory effect of verbascoside through the regulation of TAK-1/JNK/AP-1 signalling in the U937 cell line. Our results demonstrate that verbascoside increased the phosphorylation of SHP-1, by attenuating the activation of TAK-1/JNK/AP-1 signalling. This leads to a reduction in the expression and activity of both COX and NOS. Moreover, SHP-1 depletion deletes verbascoside inhibitory effects on pro-inflammatory molecules induced by LPS. Our data confirm that SHP-1 plays a critical role in restoring the physiological mechanisms of inducible proteins such as COX2 and iNOS, and that the down-regulation of TAK-1/JNK/AP-1 signalling by targeting SHP-1 should be considered as a new therapeutic strategy for the treatment of inflammatory diseases.
Asunto(s)
Regulación hacia Abajo/efectos de los fármacos , Glucósidos/farmacología , Inflamación/enzimología , Inflamación/patología , Fenoles/farmacología , Proteína Tirosina Fosfatasa no Receptora Tipo 6/metabolismo , Muerte Celular/efectos de los fármacos , Núcleo Celular/efectos de los fármacos , Núcleo Celular/metabolismo , Ciclooxigenasa 2/metabolismo , Activación Enzimática/efectos de los fármacos , Silenciador del Gen/efectos de los fármacos , Glucósidos/química , Humanos , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Quinasas Quinasa Quinasa PAM/metabolismo , Datos de Secuencia Molecular , Óxido Nítrico Sintasa de Tipo II/metabolismo , Fenoles/química , Fosforilación/efectos de los fármacos , Transducción de Señal , Factor de Transcripción AP-1/metabolismo , Tirosina/metabolismo , Células U937RESUMEN
Resistance to erythropoietin (EPO) affects a significant number of anaemic patients with end-stage renal disease. Previous reports suggest that inflammation is one of the major independent predictors of EPO resistance, and the effects of EPO treatment on inflammatory mediators are not well established. The aim of this study was to investigate EPO-induced modification to gene expression in primary cultured leucocytes. Microarray experiments were performed on primed ex vivo peripheral blood mononuclear cells (PBMCs) and treated with human EPO-α. Data suggested that EPO-α modulated genes involved in cell movement and interaction in primed PBMCs. Of note, EPO-α exerts anti-inflammatory effects inhibiting the expression of pro-inflammatory cytokine IL-8 and its receptor CXCR2; by contrast, EPO-α increases expression of genes relating to promotion of inflammation encoding for IL-1ß and CCL8, and induces de novo synthesis of IL-1α, CXCL1 and CXCL5 in primed cells. The reduction in MAPK p38-α activity is involved in modulating both IL-1ß and IL-8 expression. Unlike the induction of MAPK, Erk1/2 activity leads to upregulation of IL-1ß, but does not affect IL-8 expression and release. Furthermore, EPO-α treatment of primed cells induces the activation of caspase-1 upstream higher secretion of IL-1ß, and this process is not dependent on caspase-8 activation. In conclusion, our findings highlight new potential molecules involved in EPO resistance and confirm the anti-inflammatory role for EPO, but also suggest a plausible in vivo scenario in which the positive correlation found between EPO resistance and elevated levels of some pro-inflammatory mediators is due to treatment with EPO itself.
Asunto(s)
Eritropoyetina/metabolismo , Perfilación de la Expresión Génica , Leucocitos Mononucleares/metabolismo , Células Cultivadas , Humanos , Leucocitos Mononucleares/citologíaRESUMEN
Recently, astaxanthin (ASTA) studies have focused on several biological functions such as radical scavenging, singlet oxygen quenching, anti-carcinogenesis, anti-diabetic, anti-obesity, anti-inflammatory, anti-melanogenesis, and immune enhancement activities. In this study, we investigated the potential role protective of ASTA, an antioxidant marine carotenoid, in restoring physiological conditions in U937 cells stimulated with LPS (10 µg/ml). Our results show that pre-treatment with ASTA (10 µM) for 1 h attenuates the LPS-induced toxicity and ROS production. The beneficial effect of ASTA is associated with a reduction intracellular O2 (-) production by restoring the antioxidant network activity of superoxide dismutase (SOD) and catalase (CAT), which influence HO-1 expression and activity by inhibiting nuclear translocation of Nrf2. We accordingly hypothesize that ASTA has therapeutic properties protecting U937 cells from LPS-induced inflammatory and oxidative stress.
Asunto(s)
Lipopolisacáridos/farmacología , Estrés Oxidativo/efectos de los fármacos , Sustancias Protectoras/farmacología , Superóxidos/metabolismo , Antioxidantes/metabolismo , Catalasa/metabolismo , Supervivencia Celular/efectos de los fármacos , Hemo-Oxigenasa 1/metabolismo , Humanos , Factor 2 Relacionado con NF-E2/metabolismo , Nitroazul de Tetrazolio/metabolismo , Superóxido Dismutasa/metabolismo , Células U937 , Xantófilas/química , Xantófilas/farmacologíaRESUMEN
Analytical methods for quantification of 5'-methylcytosine in genomes are important tools to investigate epigenetic changes in gene expression during development, differentiation, aging, or cancer. Here, we report a novel genomic methylation content assay based on enzymatic hydrolysis of DNA and MEKC separation of 5'-deoxyribonucleoside monophosphates (dNMP) using the cationic surfactant CTAB as pseudostationary phase. Calf Thymus DNA was used during method development to determine electrophoretic parameters and electrolyte composition for a complete separation between 2'-deoxycytosine-5'-monophosphate and 2'-deoxy-5'-methylcytosine 5'-monophosphate (d5mCMP). Methylated and not methylated oligonucleotides were used to confirm the identity of each peak and evaluate analytical parameters of the method. The LOD of the method was found to be 12.5 pmol/µL for d5mCMP.
Asunto(s)
Cromatografía Capilar Electrocinética Micelar/métodos , Metilación de ADN , ADN/metabolismo , Espectrofotometría Ultravioleta/métodos , Animales , Bovinos , Cetrimonio , Compuestos de Cetrimonio , Citidina Monofosfato/análogos & derivados , Citidina Monofosfato/análisis , Citidina Monofosfato/química , ADN/genética , Hidrólisis , Límite de Detección , Reproducibilidad de los ResultadosRESUMEN
Recently, several studies reported a relationship between immune system activation and anger expression. Consequently, the aim of this study was to explore immunitary molecular mechanisms that potentially underlie anger expression. To this end, we applied the Frustration-Aggression Theory in a contact sport model, utilizing the nearing of sporting events to trigger anger feelings. In parallel, we evaluated the activation of immune system at mRNA levels. We enrolled 20 amateur rugby players (age ± SD, 27.2 ± 4.5) who underwent psychological assessment to evaluate anger, with the State-Trait Anger Expression Inventory-2 (STAXI-2), before rugby matches; at the same time blood samples were taken to analyze the variations of gene expression by microarray. During the 2 hr before each game, a significant increase was verified in the Rage State (RS) score compared to the score ascertained 72 hr before. At the same time, we found modulation in expression profile, in particular increased expression of gene that encodes interleukin l-ß (IL-1ß). In a regression analysis, RS score was related to IL-1ß, and the potential risk factors age, body mass index, smoking, and drinking. The levels of cytokine were positively and independently related to RS score. Our results suggest that the nearing of sporting event can trigger anger state feelings and activate immune system in rugby players. We propose the IL-1ß as a potential biological marker of anger. However, further research is necessary to clarify the correlation between cytokine and anger.
Asunto(s)
Ira/fisiología , Atletas/psicología , Fútbol Americano/fisiología , Interleucina-1beta/sangre , Adulto , Factores de Edad , Agresión/fisiología , Consumo de Bebidas Alcohólicas/psicología , Índice de Masa Corporal , Fútbol Americano/psicología , Humanos , Masculino , Factores de Riesgo , Fumar/psicología , Encuestas y CuestionariosRESUMEN
BACKGROUND: Previous reports suggest that NO may contribute to the pathophysiology of septic shock. Recently, we have synthesized and characterized a series of benzyl- and dibenzyl derivative of N-(3-aminobenzyl)acetamidine, a potent and selective inhibitor of iNOS, in vitro assay. We evaluated the molecular mechanisms by which these compounds are involved in the regulation of NOSs expression. METHODS: H9c2 cells were stimulated with lipopolysaccharide (LPS) in the presence or absence of acetamidine-derivative. The NOSs mRNA and protein, and activation of signaling pathways (Akt and NF-κB) were assayed. RESULTS: The induction of endotoxic shock in H9c2 with LPS caused an increase of inducible NOS and a down-regulation of constitutive NOS. The molecular mechanism involved in the modulation of NOSs expression in H9c2 cells upon LPS stimulation resulted in the modification of the redox state responsible for NF-kB nuclear translocation via NIK -IKKα/ß-IkBα, simultaneously to the inactivation of the PI3K/Akt pathway. The compounds acted as an anti-inflammatory modulator. CONCLUSION: These results suggest that LPS regulates the opposite NOS expression in H9c2 cells by modifying the redox state of these cells responsible for the NF-kB nuclear translocation via NIK-IKKα/ß-IkBα, simultaneous to the inactivation of the PI3K/Akt pathway. The new molecule acts as an anti-inflammatory modulator in LPS-induced inflammation in H9c2 cells by the restoration of eNOS and nNOS expressions, mechanistically involving the PI3K/Akt pathway. GENERAL SIGNIFICANCE: This study delineates the underlying mechanisms of opposite NOSs expression in H9c2 cells stimulated with LPS.
Asunto(s)
Amidinas/farmacología , Inflamación/metabolismo , Lipopolisacáridos/farmacología , Mioblastos Cardíacos/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Animales , Antiinflamatorios/farmacología , Apoptosis/efectos de los fármacos , Western Blotting , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Citometría de Flujo , Humanos , Quinasa I-kappa B/genética , Quinasa I-kappa B/metabolismo , Inflamación/inducido químicamente , Inflamación/patología , Mioblastos Cardíacos/citología , Mioblastos Cardíacos/efectos de los fármacos , FN-kappa B/genética , FN-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo II/genética , Nitritos/metabolismo , Oxidación-Reducción , Fosfatidilinositol 3-Quinasas/genética , Proteínas Proto-Oncogénicas c-akt/genética , ARN Mensajero/genética , Ratas , Especies Reactivas de Oxígeno/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transducción de Señal/efectos de los fármacosRESUMEN
In this study, the activity of the antioxidant enzyme network was assessed spectrophotometrically in samples of dental pulp and dental papilla taken from third-molar gem extracts. The production of nitric oxide by the conversion of l-(2,3,4,5)-[3H] arginine to l-(3H) citrulline, the activity of haem oxygenase 1 (HO-1) through bilirubin synthesis and the expression of inducible nitric oxide synthase (iNOS), HO-1 proteins and messenger RNA by Western blot and reverse-transcribed polymerase chain reaction were also tested. The objective of this study was to evaluate the role of two proteins, iNOS and HO-1, which are upregulated by a condition of oxidative stress present during dental tissue differentiation and development. This is fundamental for guaranteeing proper homeostasis favouring a physiological tissue growth. The results revealed an over-expression of iNOS and HO-1 in the papilla, compared with that in the pulp, mediated by the nuclear factor kappa B transcription factor activated by the reactive oxygen species that acts as scavengers for the superoxide radicals. HO-1, a metabolically active enzyme in the papilla, but not in the pulp, seems to inhibit the iNOS enzyme by a crosstalk between the two proteins. We suggest that the probable mechanism through which this happens is the interaction of HO-1 with haem, a cofactor dimer indispensible for iNOS, and the subsequent suppression of its metabolic activity.
Asunto(s)
Papila Dental/enzimología , Papila Dental/crecimiento & desarrollo , Cavidad Pulpar/enzimología , Cavidad Pulpar/crecimiento & desarrollo , Regulación del Desarrollo de la Expresión Génica , Hemo-Oxigenasa 1/genética , Óxido Nítrico Sintasa de Tipo II/genética , Adolescente , Niño , Femenino , Regulación Enzimológica de la Expresión Génica , Hemo-Oxigenasa 1/metabolismo , Humanos , Masculino , FN-kappa B/genética , FN-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Regulación hacia ArribaRESUMEN
Matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) are the main determinants of tissue remodeling in both physiological and pathological processes. Metabolic processes, which generate oxidants and antioxidants can be influenced by environmental factors such as electromagnetic fields (EMF). We analyzed the effects of EMF on the activity and expression of MMPs in THP-1 cells. Cells were exposed to a 50 Hz, 1 mT EMF for 24 h and incubated with or without LPS. Our data indicate that THP-1 cells exposed to EMF causes a reduction of anti-oxidant enzyme activity and an enhancement of nitrogen intermediates involving the iNOS pathway. We then analyzed the role of nitration of TIMP-1 in increasing the activity of MMPs in EMF exposed cells. Molecular modeling tools were employed to identify the most plausible sites in the active conformation of TIMP-1; at least two protein sites, Y120 and Y38 and/or Y72 were identified. Reactive nitrogen species (RNS) may affect protein targets, such as TIMP-1, which are crucial for the regulation of MMP activities by oxidation of sulfydryl groups, or by nitration of tyrosine residues. These results may suggest a pathway connecting an imbalance of MMPs and their cognate inhibitor TIMP-1.
Asunto(s)
Campos Electromagnéticos , Leucemia Mieloide Aguda/enzimología , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Inhibidor Tisular de Metaloproteinasa-1/metabolismo , Antioxidantes/metabolismo , Línea Celular Tumoral , Regulación Enzimológica de la Expresión Génica , Humanos , Lipopolisacáridos/farmacología , Metaloproteinasa 2 de la Matriz/genética , Metaloproteinasa 9 de la Matriz/genética , Modelos Moleculares , Óxido Nítrico Sintasa de Tipo II/metabolismo , Procesamiento Proteico-Postraduccional , ARN Mensajero/metabolismo , Especies de Nitrógeno Reactivo/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Factores de Tiempo , Inhibidor Tisular de Metaloproteinasa-1/química , Inhibidor Tisular de Metaloproteinasa-1/genética , TirosinaRESUMEN
Verbascum thapsus commonly known as 'mullein' is part of a large family of Scrophulariaceae consisting of more than 360 species. From antiquity Verbascum thapsus has been used as a medicinal herb, it contains diverse polysaccharides, iroid glycosides, flavonoids, saponins, volatile oils and phenylentanoids. Inducible nitric oxide synthase (iNOS) represents one of the three isoforms that produce nitric oxide using L-arginine as a substrate in response to an increase in superoxide anion activated by NF-kB. It is implicated in different pathophysiological events and its expression increases greatly during an inflammatory process, due to oxidative stress and the activation of the enzymes of the antioxidant network such as SOD, CAT and GPx.In this study an inflammatory state was reproduced by treating THP-1 cells (human myelomonocytic leukaemia) with pro-inflammatory stimuli, such as LPS and IFN-gamma, obtaining an up-regulation both in the expression and in the activity of iNOS. The aim of the work was to investigate the antiinflammatory action of verbascoside using a concentration of 100 mum. The results show a significant decrease of the expression and activity of iNOS, extracellular O(2) (-) production, SOD, CAT and GPx activity when the cells were treated with verbascoside. Based on these results it is hypothesized that verbascoside has antiinflammatory properties since it reduces the production of superoxide radicals and consequently reduces the activity of iNOS.
Asunto(s)
Antiinflamatorios/farmacología , Antioxidantes/metabolismo , Glucósidos/farmacología , Inflamación/tratamiento farmacológico , Óxido Nítrico Sintasa de Tipo II/metabolismo , Fenoles/farmacología , Extractos Vegetales/farmacología , Verbascum/química , Antiinflamatorios/uso terapéutico , Antioxidantes/farmacología , Antioxidantes/uso terapéutico , Catalasa/metabolismo , Línea Celular Tumoral , Radicales Libres/metabolismo , Glucósidos/uso terapéutico , Glutatión Peroxidasa/metabolismo , Humanos , Inflamación/inducido químicamente , Inflamación/metabolismo , Interferón gamma , Leucemia Mieloide , Lipopolisacáridos , Estrés Oxidativo/efectos de los fármacos , Oxígeno/metabolismo , Fenoles/uso terapéutico , Componentes Aéreos de las Plantas , Extractos Vegetales/uso terapéutico , Superóxido Dismutasa/metabolismo , Regulación hacia ArribaRESUMEN
The growth of malignant tumors is determined in large part by the proliferative capacity of the tumor cells. Clinical observations and animal experiments have established that tumor cells elicit immune responses. Histopathologic studies show that many tumors are surrounded by mononuclear cell and mast cell infiltrates. Mast cells are ubiquitous in the body and are critical for allergic reactions. Increasing evidence indicates that mast cells secrete proinflammatory cytokines and are involved in neuro-inflammatory processes and cancer. Mast cells accumulate in the stroma surrounding certain tumors, especially mammary adenocarcinoma, and the molecules they secrete can benefit the tumor. However, mast cells can also increase at the site of tumor growth and participate in tumor rejection. Mast cells may be recruited by tumor-derived chemoattractants and selectively secrete molecules such as growth factors, histamine, heparin, VEGF, and IL-8, as well as proteases that permit the formation of new blood vessels and metastases. Tumor mast cell intersections play regulatory and modulatory roles affecting various aspects of tumor growth. Discovery of these new roles of mast cells further complicates the understanding of tumor growth. This review focuses on the strategic importance of mast cells to the progression of tumors, and proposes a revised immune effector mechanism of mast cell involvement in tumor growth.
Asunto(s)
Mastocitos/inmunología , Neoplasias/fisiopatología , Proliferación Celular , Citocinas/metabolismo , Histamina/metabolismo , Humanos , Mastocitos/metabolismo , Neoplasias/inmunología , Neovascularización Patológica/inmunologíaRESUMEN
The purpose of this study was to investigate whether overnight exposure to 1 mT-50 Hz extremely low-frequency sinusoidal electromagnetic field (EMF) affects the expression and production of inducible nitric oxide synthase (iNOS) and monocyte chemotactic protein-1 (MCP-1) in human monocytes. RT-PCR and Western blot analysis demonstrate that EMF exposure affects the expression of iNOS and MCP-1 in cultured human mononuclear cells at the mRNA level and protein synthesis. Interestingly, the effects of EMF exposure clearly differed with respect to the potentiation and inhibition of iNOS and MCP-1 expression. Whereas iNOS was down-regulated both at the mRNA level and at the protein level, MCP-1 was up-regulated. These results provide helpful information regarding the EMF-mediated modulation of the inflammatory response in vivo. However, additional studies are necessary to demonstrate that EMF acts as a nonpharmacological inhibitor of NO and inducer of MCP-1 in some diseases where the balance of MCP-1 and NO may be important.
Asunto(s)
Quimiocina CCL2/efectos de la radiación , Campos Electromagnéticos , Monocitos/efectos de la radiación , Óxido Nítrico Sintasa de Tipo II/efectos de la radiación , Quimiocina CCL2/genética , Femenino , Expresión Génica/efectos de la radiación , Humanos , Lipopolisacáridos/inmunología , Masculino , Persona de Mediana Edad , Monocitos/enzimología , Monocitos/inmunología , Monocitos/metabolismo , FN-kappa B/metabolismo , FN-kappa B/efectos de la radiación , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Biosíntesis de Proteínas/efectos de la radiación , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
OBJECTIVE: Due to reported modulatory effects of statins on nitric oxide synthase (NOS) expression, we tested the hypothesis of protective effects of in vivo chronic treatment with rosuvastatin, a novel 3-hydroxy-3-methyl-glutaryl coenzyme A-reductase inhibitor, on ischemia-reperfusion injury, and investigated mechanisms involved. METHODS: After 3 weeks of in vivo treatment with rosuvastatin (0.2-20 mg/kg/day) or placebo, excised hearts from Wistar rats were subjected to 15 min global ischemia and 22-180 min reperfusion. We evaluated creatine-phosphokinase and nitrite levels in the coronary effluent, heart weight changes, microvascular permeability (extravasation of fluoresceine-labeled albumin), ultrastructural alterations, and the expression of endothelial (e) and inducible (i) nitric oxide synthase (NOS) (by reverse-transcription polymerase chain reaction and Western blotting). RESULTS: Rosuvastatin 0.2 and 2 mg/kg/day significantly reduced myocardial damage and vascular hyperpermeability, concomitant with a reduction in endothelial and cardiomyocyte lesions. At 2 mg/kg/day, rosuvastatin significantly increased eNOS mRNA and protein compared with untreated hearts, and conversely decreased iNOS mRNA and protein, as well as nitrite production after ischemia-reperfusion. The addition of the NOS inhibitor N(omega)-nitro-L-arginine methylester (L-NAME, 30 micromol/L) significantly reduced cardioprotection against ischemia-reperfusion. CONCLUSIONS: Chronic treatment with rosuvastatin before ischemia reduces ischemia-reperfusion injury and prevents coronary endothelial cell and cardiomyocyte damage by NO-dependent mechanisms.
Asunto(s)
Fluorobencenos/uso terapéutico , Inhibidores de Hidroximetilglutaril-CoA Reductasas/uso terapéutico , Daño por Reperfusión Miocárdica/tratamiento farmacológico , Óxido Nítrico Sintasa/metabolismo , Pirimidinas/uso terapéutico , Sulfonamidas/uso terapéutico , Animales , Western Blotting/métodos , Permeabilidad Capilar , Creatina Quinasa/análisis , Esquema de Medicación , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/patología , Miocardio/metabolismo , Miocardio/ultraestructura , Óxido Nítrico Sintasa/genética , Nitritos/análisis , Perfusión , ARN Mensajero/análisis , Ratas , Ratas Wistar , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Rosuvastatina CálcicaRESUMEN
BACKGROUND: High prevalence of atherosclerotic cardiovascular events accounts for much of the mortality among patients suffering from end-stage renal disease (ESRD). Endothelial dysfunction as a pathogenic mechanism might contribute to increasing the cardiovascular risk of ESRD. Reduced endothelium-dependent vasodilation has consistently been observed in chronic renal failure patients. Since nitric oxide (NO) is the principal endothelium-derived vasodilator, a reduction in the NO bioavailability may be envisaged in ESRD patients. METHODS: To clarify whether exposure to erythrocytes from ESRD patients might modulate NO release by the endothelium, we evaluated endothelial NO synthase (eNOS) protein levels (Western blot), eNOS mRNA quantity (real-time PCR), and NOS activity (conversion of L-[3H] arginine in L-[3H] citruline) in endothelial cultures stimulated by erythrocytes from healthy subjects and ESRD patients. RESULTS: A time-dependent decrease in eNOS protein levels was evident in cultures treated with erythrocytes from ESRD patients. This observation was consistent with the decreased eNOS mRNA quantities induced by erythrocytes from such patients. Moreover, compared to controls, NOS activity exhibited a significant reduction after incubation with erythrocytes from ESRD patients. The observed eNOS reduction induced by erytrocytes from ESRD patients was totally abolished by annexin V, able to mask red blood cell (RBC) surface-exposed phosphatidylserine. CONCLUSION: These findings suggest that adhesion of erythrocytes from ESRD patients to vascular endothelium may cause a decrease in the levels of eNOS mRNA and protein, and inhibition of NOS activity. This might contribute to endothelial dysfunction, and may play a role in the pathogenesis of cardiovascular disease in ESRD patients.
Asunto(s)
Adhesión Celular/fisiología , Endotelio Vascular/enzimología , Eritrocitos/fisiología , Óxido Nítrico Sintasa/metabolismo , Uremia/sangre , Uremia/enzimología , Anciano , Enfermedades Cardiovasculares/etiología , Estudios de Casos y Controles , Células Cultivadas , Endotelio Vascular/citología , Eritrocitos/patología , Femenino , Expresión Génica , Humanos , Técnicas In Vitro , Fallo Renal Crónico/sangre , Fallo Renal Crónico/complicaciones , Fallo Renal Crónico/enzimología , Masculino , Persona de Mediana Edad , Óxido Nítrico Sintasa/genética , Óxido Nítrico Sintasa de Tipo III , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
Interleukin-17 (now known as IL-17A), is a homodimer of two 155 amino acid chains secreted by CD4+ activated memory T cells (CD45+ RO+) and is available as a glycosylated 20- to 30-kDa homodimeric peptide. Human IL-17 shows amino acid sequence identity of 62.5 and 58% to the mouse and rat sequences, respectively. IL-17 can regulate the function of a variety of cell types, plays an important role in the maturation of hematopoietic progenitor cells, and induces production of proinflammatory mediators. Here, for the first time, we summarize the biological effects of IL-17 and its family members as important players of T cell-mediated immune responses and underline the important implications of this cytokine in inflammation and degenerative diseases.