RESUMEN
AIM: To investigate the effect of (-)-epigallocatechin-3-gallate (EGCG) on polyinosinic-polycytidylic acid (poly I:C)-triggered intracellular innate immunity against hepatitis C virus (HCV) in hepatocytes. METHODS: A cell culture model of HCV infection was generated by infecting a hepatoma cell line, Huh7, with HCV JFH-1 strain (JFH-1-Huh7). Poly I:C with a high molecular weight and EGCG were used to stimulate the JFH-1-Huh7 cells. Real-time reverse transcription-polymerase chain reaction was used to detect the expression levels of intracellular mRNAs and of intracellular and extracellular HCV RNA. Enzyme-linked immunosorbent assay was used to evaluate the interferon (IFN)-λ1 protein level in the cell culture supernatant. Immunostaining was used to examine HCV core protein expression in Huh7 cells. RESULTS: Our recent study showed that HCV replication could impair poly I:C-triggered intracellular innate immune responses in hepatocytes. In the current study, we showed that EGCG treatment significantly increased the poly I:C-induced expression of Toll-like receptor 3 (TLR3), retinoic acid-inducible gene I, and IFN-λ1 in JFH-1-Huh7 cells. In addition, supplementation with EGCG increased the poly I:C-mediated antiviral activity in JFH-1-Huh7 cells at the intracellular and extracellular HCV RNA and protein levels. Further investigation of the mechanisms showed that EGCG treatment significantly enhanced the poly I:C-induced expression of IFN-regulatory factor 9 and several antiviral IFN-stimulated genes, including ISG15, ISG56, myxovirus resistance A, and 2'-5'-oligoadenylate synthetase 1, which encode the key antiviral elements in the IFN signaling pathway. CONCLUSION: Our observations provide experimental evidence that EGCG has the ability to enhance poly I:C-induced intracellular antiviral innate immunity against HCV replication in hepatocytes.
Asunto(s)
Antivirales/farmacología , Catequina/análogos & derivados , Hepacivirus/fisiología , Hepatitis C/inmunología , Inmunidad Innata/efectos de los fármacos , Interferón gamma/inmunología , Poli I-C/inmunología , Antivirales/uso terapéutico , Catequina/farmacología , Catequina/uso terapéutico , Línea Celular Tumoral , Proteína 58 DEAD Box/inmunología , Proteína 58 DEAD Box/metabolismo , Ensayo de Inmunoadsorción Enzimática , Hepatitis C/tratamiento farmacológico , Hepatitis C/virología , Hepatocitos , Humanos , Subunidad gamma del Factor 3 de Genes Estimulados por el Interferón/inmunología , Subunidad gamma del Factor 3 de Genes Estimulados por el Interferón/metabolismo , Interferón gamma/metabolismo , ARN Viral/aislamiento & purificación , Receptores Inmunológicos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transducción de Señal/inmunología , Receptor Toll-Like 3/metabolismo , Proteínas del Núcleo Viral/metabolismo , Replicación Viral/efectos de los fármacos , Replicación Viral/inmunologíaRESUMEN
Lipopolysaccharide- (LPS-) mediated systemic inflammation plays a critical role in neurodegenerative diseases. The present study was conducted to evaluate the protective effects of epigallocatechin gallate (EGCG), the major component in green tea, on LPS-mediated inflammation and neurotoxicity. LPS treatment of macrophages induced expression of proinflammatory cytokines (TNF-α, IL-1ß, and IL-6). However, EGCG pretreatment of macrophages significantly inhibited LPS-mediated induction of these cytokines. In addition, EGCG significantly diminished LPS-induced inflammatory cytokines in the peripheral mononuclear blood cells (PBMCs). Supernatant from EGCG-pretreated and LPS-activated macrophage cultures was found to be less cytotoxic to neurons than that from non-EGCG-pretreated and LPS-activated macrophage cultures. Furthermore, EGCG treatment of neurons could inhibit LPS-induced production of reactive oxygen species (ROS). Thus EGCG represents a potent and useful neuroprotective agent for inflammation-mediated neurological disorders.
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Catequina/análogos & derivados , Inflamación/prevención & control , Leucocitos Mononucleares/inmunología , Lipopolisacáridos/inmunología , Macrófagos/inmunología , Neuronas/fisiología , Fármacos Neuroprotectores/farmacología , Animales , Catequina/química , Catequina/farmacología , Supervivencia Celular , Células Cultivadas , Medios de Cultivo/química , Humanos , Inflamación/terapia , Mediadores de Inflamación/inmunología , Interleucina-1beta/genética , Interleucina-6/genética , Leucocitos Mononucleares/efectos de los fármacos , Lipopolisacáridos/administración & dosificación , Activación de Macrófagos , Macrófagos/efectos de los fármacos , Neuronas/efectos de los fármacos , Neuronas/inmunología , Fármacos Neuroprotectores/química , Ratas , Especies Reactivas de Oxígeno/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Té/química , Factor de Necrosis Tumoral alfa/genéticaRESUMEN
PURPOSE: Docosahexaenoic acid (DHA) was supplemented in a single-site, placebo-controlled, randomized clinical trial designed to slow vision loss associated with X-linked retinitis pigmentosa (XLRP); the DHAX Trial. We previously reported no significant differences between supplemented and placebo groups in intent-to-treat analysis of primary ERG outcomes. Assessed herein are hypothesis-generating measures of ancillary visual function outcomes in participants fully adhering to trial protocol. METHODS: Male participants with XLRP (range, 7-31 years) received 30 mg DHA/kg/d (n = 29) or placebo (n = 22) for 4 years. Visual outcomes were measured annually and red blood cell (RBC) DHA determined every 6 months. RESULTS: Oral DHA supplementation increased mean RBC-DHA levels by 4-fold (P < 0.0001) over placebo. No group differences in progression were found for visual acuity (P = 0.11), shape discrimination (P = 0.18), or fundus appearance (P = 0.70). Optical coherence tomography (OCT) became available during year 2 of the trial; no group differences were seen in ellipsoid zone constriction (P = 0.87) over 2 years. Yearly rates of progression were reduced for dark-adapted thresholds (P = 0.06) and visual field sensitivity for foveal, macular, peripheral, total, and ellipsoid zone regions by DHA supplementation (P = 0.039, P = 0.031, P < 0.0001, P < 0.0001, and P = 0.033). Rates of visual field sensitivity decline were dependent on RBC-DHA (P = 0.046 to <0.0001). CONCLUSIONS: Supplementation of DHA significantly elevated blood DHA levels and reduced the rate of progression in final dark-adapted thresholds and visual field sensitivity. From the relationship between RBC-DHA and the rate of field sensitivity loss, we can extrapolate that an RBC-DHA level of 17% could minimize the decline in field sensitivity. (ClinicalTrials.gov number, NCT00100230.)
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Ácidos Docosahexaenoicos/uso terapéutico , Enfermedades Genéticas Ligadas al Cromosoma X/tratamiento farmacológico , Retinitis Pigmentosa/tratamiento farmacológico , Adolescente , Adulto , Niño , Progresión de la Enfermedad , Percepción de Forma/efectos de los fármacos , Fondo de Ojo , Humanos , Masculino , Retinitis Pigmentosa/genética , Campos Visuales/efectos de los fármacos , Adulto JovenRESUMEN
Berberine, which is a wellknown drug used in traditional medicine, has been demonstrated to exert diverse pharmacological effects, including antiinflammatory effects. However, whether berberine can affect the production of inflammatory molecules in vascular endothelial cells remains to be elucidated. Therefore, the present study aimed to determine the effects of berberine, and the underlying molecular mechanisms of these effects. The effect of berberine on tumor necrosis factor (TNF)αinduced inflammatory molecule expression was examined in cultured human aortic endothelial cells (HAECs). The HAECs were stimulated with TNFα and incubated with or without berberine. The activation of nuclear factor (NF)κB and adenosine monophosphateactivated protein kinase (AMPK) were analyzed using western blotting, and the protein secretion of intercellular adhesion molecule (ICAM)1 and monocyte chemoattractant protein (MCP)1 was measured using ELISA kits. The mRNA expression levels of ICAM1 and MCP1 were analyzed using reverse transcriptionquantitative polymerase chain reaction. The results of the present study demonstrated that berberine significantly inhibited the TNFαinduced expression of ICAM1 and MCP1, as well as the activation of NFκB in the HAECs. These effects were attenuated following cotreatment with AMPK inhibitor compound C, or specific small interfering RNAs. In conclusion, the results of the present study indicated that berberine inhibits the TNFαinduced expression of ICAM1 and MCP1, and the activation of NFκB in HAECs in vitro, possibly through the AMPKdependent pathway.
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Proteínas Quinasas Activadas por AMP/genética , Antiinflamatorios no Esteroideos/farmacología , Berberina/farmacología , Células Endoteliales/efectos de los fármacos , FN-kappa B/genética , Factor de Necrosis Tumoral alfa/farmacología , Proteínas Quinasas Activadas por AMP/antagonistas & inhibidores , Proteínas Quinasas Activadas por AMP/metabolismo , Línea Celular , Quimiocina CCL2/agonistas , Quimiocina CCL2/antagonistas & inhibidores , Quimiocina CCL2/genética , Quimiocina CCL2/metabolismo , Células Endoteliales/citología , Células Endoteliales/metabolismo , Regulación de la Expresión Génica , Humanos , Inflamación , Molécula 1 de Adhesión Intercelular/genética , Molécula 1 de Adhesión Intercelular/metabolismo , FN-kappa B/agonistas , FN-kappa B/antagonistas & inhibidores , FN-kappa B/metabolismo , Inhibidores de Proteínas Quinasas/farmacología , ARN Mensajero/agonistas , ARN Mensajero/antagonistas & inhibidores , ARN Mensajero/genética , ARN Mensajero/metabolismo , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Transducción de Señal , Factor de Necrosis Tumoral alfa/antagonistas & inhibidoresRESUMEN
This study is to observe allergic response to Qingkailing injection in BN rats and to establish a suitable animal model to evaluate allergic response induced by traditional Chinese medicine. BN rats were sensitized by Qingkailing injection, and guinea pigs were similarly sensitized as the control. The symptoms of allergic response were observed, the levels of histamine in serum and tissues were determined by ELISA assay and pathological changes in lung and trachea were observed with HE staining under light microscope. The total incidence of allergic response in BN rats was 52.78%, which was higher than that in guinea pig groups (16.67%). The total degree of allergic response in BN rats was higher than that in guinea pigs. Compared with control groups, the level of histamine in serum, lung and trachea tissues of BN rats and guinea pigs increased significantly. The release rate of histamine in BN rats was higher than that in guinea pigs. The rate and degree of pathological changes in lung and trachea tissues of BN rats were higher than that in guinea pigs. Compared with guinea pig, BN rat is probably a suitable animal model in evaluating allergic response to injection of traditional Chinese medicine.
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Modelos Animales de Enfermedad , Medicamentos Herbarios Chinos/efectos adversos , Hipersensibilidad Inmediata/inducido químicamente , Animales , Cobayas , Histamina/sangre , Inyecciones , Masculino , Ratas , Ratas Endogámicas BNRESUMEN
OBJECTIVE: To observe the effect of the extract from gardenia on influenza viral pneumonia in mice and virus-induced cytopathic effect. METHOD: The mice were infected by influenza virus in nasal, the lung inflammation, mortality rate and life elongation rate were observed respectively. The anti-viral activity of the extract from gardenia was accessed by cytopathic effect (CPE) in vitro and 0% toxicity concentration (TC0), 50% toxicity concentration (TC50), 50% inhibitor concentration (IC50), therapeutic index (TI) were determined by Reed-Muench method. RESULT: The pneumonia induced by influenza virus in mice was inhibited significantly by the extract from gardenia, as the mortality rate decreased and the life elongation rate increased remarkably. Meanwhile the NO content in serum decreased significantly; The cytopathic effect induced by six kinds of viruses was inhibited remarkably. CONCLUSION: The six kinds of viruses were inhibited significantly by the extract from gardenia which inhibitory effect on mice influenza viral pneumonia was related to the NO content decreased.