RESUMEN
Osteoarthritis (OA) is characterized by the degeneration and destruction of articular cartilage. Allicin, a dietary garlic active constituent, exerts anti-inflammatory effects on several diseases. However, its effects on OA have not been clearly elucidated. In this study, we explored the effects of allicin on OA in both in vitro and in vivo models. Allicin inhibited interleukin-1ß (IL-1ß) induced overproduction of nitric oxide, inducible nitric oxide synthase, prostaglandin E2, and cyclooxygenase-2, as well as pro-inflammatory cytokines tumor necrosis factor alpha and interleukin-6 in chondrocytes in a dose-dependent manner. Meanwhile, allicin reversed the overproduction of metalloproteinase-13 and a disintegrin and metalloproteinase with thrombospondin motifs-5 and the decrease of aggrecan and type II collagen. Furthermore, allicin dramatically suppressed IL-1ß-stimulated PI3K/Akt/NF-κB activation in chondrocytes. In vivo, treatment with allicin prevented the destruction of cartilage and inhibited PI3K/Akt/NF-κB activation in the cartilage of mice OA models. Taken together, these results indicate that allicin may be a potential therapeutic agent for OA.
Asunto(s)
Conservadores de la Densidad Ósea/uso terapéutico , Condrocitos/metabolismo , Suplementos Dietéticos , Modelos Animales de Enfermedad , Osteoartritis de la Rodilla/terapia , Inhibidores de las Quinasa Fosfoinosítidos-3 , Ácidos Sulfínicos/uso terapéutico , Animales , Conservadores de la Densidad Ósea/efectos adversos , Conservadores de la Densidad Ósea/metabolismo , Supervivencia Celular , Células Cultivadas , Condrocitos/citología , Condrocitos/patología , Suplementos Dietéticos/efectos adversos , Progresión de la Enfermedad , Disulfuros , Femenino , Regulación de la Expresión Génica , Humanos , Interleucina-1beta/antagonistas & inhibidores , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Masculino , Ratones Endogámicos C57BL , FN-kappa B/agonistas , FN-kappa B/antagonistas & inhibidores , FN-kappa B/metabolismo , Osteoartritis de la Rodilla/metabolismo , Osteoartritis de la Rodilla/patología , Osteoartritis de la Rodilla/fisiopatología , Fosfatidilinositol 3-Quinasa/química , Fosfatidilinositol 3-Quinasa/metabolismo , Proteínas Proto-Oncogénicas c-akt/agonistas , Proteínas Proto-Oncogénicas c-akt/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-akt/metabolismo , Distribución Aleatoria , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Transducción de Señal , Ácidos Sulfínicos/efectos adversos , Ácidos Sulfínicos/metabolismoRESUMEN
UNLABELLED: Chemical regulation of macrophage function is one key strategy for developing host-directed adjuvant therapies for tuberculosis (TB). A critical step to develop these therapies is the identification and characterization of specific macrophage molecules and pathways with a high potential to serve as drug targets. Using a barcoded lentivirus-based pooled short-hairpin RNA (shRNA) library combined with next generation sequencing, we identified 205 silenced host genes highly enriched in mycobacteria-resistant macrophages. Twenty-one of these "hits" belonged to the oxidoreductase functional category. NAD(P)H: quinone oxidoreductase 1 (NQO1) was the top oxidoreductase "hit". NQO1 expression was increased after mycobacterial infection, and NQO1 knockdown increased macrophage differentiation, NF-κB activation, and the secretion of pro-inflammatory cytokines TNF-α and IL-1ß in response to infection. This suggests that mycobacteria hijacks NQO1 to down-regulate pro-inflammatory and anti-bacterial functions. The competitive inhibitor of NQO1 dicoumarol synergized with rifampin to promote intracellular killing of mycobacteria. Thus, NQO1 is a new host target in mycobacterial infection that could potentially be exploited to increase antibiotic efficacy in vivo. Our findings also suggest that pooled shRNA libraries could be valuable tools for genome-wide screening in the search for novel druggable host targets for adjunctive TB therapies.
Asunto(s)
Antituberculosos/farmacología , Dicumarol/farmacología , Interacciones Huésped-Patógeno/efectos de los fármacos , Macrófagos/inmunología , Mycobacterium tuberculosis/efectos de los fármacos , NAD(P)H Deshidrogenasa (Quinona)/genética , Sinergismo Farmacológico , Inhibidores Enzimáticos/farmacología , Regulación de la Expresión Génica , Biblioteca de Genes , Secuenciación de Nucleótidos de Alto Rendimiento , Ensayos Analíticos de Alto Rendimiento , Humanos , Interleucina-1beta/agonistas , Interleucina-1beta/genética , Interleucina-1beta/inmunología , Macrófagos/efectos de los fármacos , Macrófagos/microbiología , Mycobacterium tuberculosis/patogenicidad , Mycobacterium tuberculosis/fisiología , NAD(P)H Deshidrogenasa (Quinona)/antagonistas & inhibidores , NAD(P)H Deshidrogenasa (Quinona)/inmunología , FN-kappa B/agonistas , FN-kappa B/genética , FN-kappa B/inmunología , Cultivo Primario de Células , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Rifampin/farmacología , Transducción de Señal , Células THP-1 , Factor de Necrosis Tumoral alfa/agonistas , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
The compound 5-fluorouracil (5-FU) is used in cancer chemotherapy and is known to cause diarrhoea. We recently reported that chemokine (C-X-C motif) ligand 1 (CXCL1) and neutrophils in the colonic mucosa were markedly increased by the administration of 5-FU in mice. Curcumin has anti-inflammatory, antitumour and antioxidant properties. Therefore, we examined the effect of curcumin on 5-FU-induced diarrhoea development and CXCL1 and CXCL2 up-regulation in the colon. Mice were given 5-FU (50 mg/kg, i.p.) daily for 4 days. Curcumin (100 or 300 mg/kg, p.o.) was administered on the day before the first administration of 5-FU and administered 30 min. before the administration of 5-FU. Gene expression levels of CXCL1 and CXCL2 in the colon were examined by real-time RT-PCR. Curcumin reduced the 5-FU-induced diarrhoea development. Under this condition, the CXCL1 and CXCL2 gene up-regulated by 5-FU administration was inhibited by curcumin. The gene expression of CXCL1 and CXCL2 was also enhanced by 5-FU application in vitro. The 5-FU-induced up-regulated CXCL1 and CXCL2 gene expressions were inhibited by curcumin, Bay-117082 and bortezomib, nuclear factor kappa B (NF-κB) inhibitors, C646, a p300/cyclic adenosine monophosphate response element-binding protein-histone acetyltransferase (HAT) inhibitor. In conclusion, these findings suggested that curcumin prevented the development of diarrhoea by inhibiting NF-κB and HAT activation.
Asunto(s)
Antimetabolitos Antineoplásicos/efectos adversos , Antioxidantes/uso terapéutico , Colon Descendente/efectos de los fármacos , Curcumina/uso terapéutico , Diarrea/prevención & control , Suplementos Dietéticos , Fluorouracilo/efectos adversos , Animales , Antiinflamatorios no Esteroideos/administración & dosificación , Antiinflamatorios no Esteroideos/farmacología , Antiinflamatorios no Esteroideos/uso terapéutico , Antimetabolitos Antineoplásicos/química , Antimetabolitos Antineoplásicos/farmacología , Antioxidantes/administración & dosificación , Antioxidantes/farmacología , Quimiocina CXCL1/agonistas , Quimiocina CXCL1/antagonistas & inhibidores , Quimiocina CXCL1/genética , Quimiocina CXCL1/metabolismo , Quimiocina CXCL2/agonistas , Quimiocina CXCL2/antagonistas & inhibidores , Quimiocina CXCL2/genética , Quimiocina CXCL2/metabolismo , Colon Descendente/inmunología , Colon Descendente/metabolismo , Colon Descendente/fisiopatología , Curcumina/administración & dosificación , Curcumina/farmacología , Diarrea/inducido químicamente , Diarrea/metabolismo , Diarrea/fisiopatología , Proteína p300 Asociada a E1A/agonistas , Proteína p300 Asociada a E1A/antagonistas & inhibidores , Proteína p300 Asociada a E1A/metabolismo , Inhibidores Enzimáticos/farmacología , Fluorouracilo/antagonistas & inhibidores , Fluorouracilo/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Mucosa Intestinal/efectos de los fármacos , Mucosa Intestinal/inmunología , Mucosa Intestinal/metabolismo , Mucosa Intestinal/fisiopatología , Masculino , Ratones Endogámicos C57BL , FN-kappa B/agonistas , FN-kappa B/antagonistas & inhibidores , FN-kappa B/metabolismo , Índice de Severidad de la Enfermedad , Técnicas de Cultivo de TejidosRESUMEN
We herein report the immunostimulatory effect of spinach aqueous extract (SAE) on mouse macrophage-like J774.1 cells and mouse primary peritoneal macrophages. SAE significantly enhanced the production of interleukin (IL)-6 and tumor necrosis factor-α by both J774.1 cells and peritoneal macrophages by enhancing the expression levels of these cytokine genes. In addition, the phagocytosis activity of J774.1 cells was facilitated by SAE. Immunoblot analysis revealed that SAE activates mitogen-activated protein kinase and nuclear factor-κB cascades. It was found that SAE activates macrophages through not only TLR4, but also other receptors. The production of IL-6 was significantly enhanced by peritoneal macrophages from SAE-administered BALB/c mice, suggesting that SAE has a potential to stimulate macrophage activity in vivo. Taken together, these data indicate that SAE would be a beneficial functional food with immunostimulatory effects on macrophages.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Interleucina-6/agonistas , Macrófagos Peritoneales/efectos de los fármacos , Extractos Vegetales/farmacología , Spinacia oleracea/química , Factor de Necrosis Tumoral alfa/agonistas , Animales , Línea Celular , Femenino , Expresión Génica , Interleucina-6/genética , Interleucina-6/inmunología , Macrófagos Peritoneales/citología , Macrófagos Peritoneales/inmunología , Ratones , Ratones Endogámicos BALB C , Proteínas Quinasas Activadas por Mitógenos/genética , Proteínas Quinasas Activadas por Mitógenos/inmunología , FN-kappa B/agonistas , FN-kappa B/genética , FN-kappa B/inmunología , Fagocitosis/efectos de los fármacos , Cultivo Primario de Células , ARN Mensajero/agonistas , ARN Mensajero/genética , ARN Mensajero/inmunología , Receptor Toll-Like 4/genética , Receptor Toll-Like 4/inmunología , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
Lupeol exhibits anti-inflammatory effects; unfortunately it shows low water solubility. An alternative to overcome this is the development of nanomaterials. Several methods for nanomaterial production are available. One of them is emulsification/solvent-evaporation. The objective of the present work was to evaluate physical properties, transport and in vitro modulator effects on NF-κB of poly (lactide-co-glycolide) (PLGA) nanoparticles loaded with lupeol. Nanonutraceuticals were prepared with 16% (w/v) of lupeol. Size distribution and morphology were measured by particle size analyzer and TEM. In vitro release of lupeol was studied by three different models: Higuchi, Siepmann & Peppas, and Power law. Transport of nanonutraceutical was studied in a Caco-2 cell model and by GC-MS. Modulator effect on NK-κB was studied by western blot analysis. Nanonutraceuticals were 10% larger than the nanoparticles without lupeol (372 vs 337 nm) and presented a broader size distribution (0.28 vs 0.22). TEM results displayed spherical structures with a broader size distribution. Entrapment efficiency of lupeol was 64.54% and it in vitro release data fitted well to the Power law and Higuchi equation (R > 0.84-0.84). Strong regulation of NF-κB of nanonutraceutical was observed. It was not observed any transport across the Caco-2 cell model at the different experimental conditions.
Asunto(s)
Antiinflamatorios no Esteroideos/metabolismo , Enterocitos/metabolismo , Absorción Intestinal , FN-kappa B/metabolismo , Nanopartículas/efectos adversos , Triterpenos Pentacíclicos/metabolismo , Poliglactina 910/efectos adversos , Transporte Activo de Núcleo Celular , Algoritmos , Antiinflamatorios no Esteroideos/administración & dosificación , Antiinflamatorios no Esteroideos/análisis , Antiinflamatorios no Esteroideos/química , Células CACO-2 , Permeabilidad de la Membrana Celular , Supervivencia Celular , Fenómenos Químicos , Suplementos Dietéticos/análisis , Emulsiones , Humanos , Microscopía Electrónica de Transmisión , FN-kappa B/agonistas , FN-kappa B/antagonistas & inhibidores , Nanopartículas/química , Nanopartículas/ultraestructura , Tamaño de la Partícula , Triterpenos Pentacíclicos/administración & dosificación , Triterpenos Pentacíclicos/análisis , Triterpenos Pentacíclicos/química , Poliglactina 910/química , Polivinilos/química , Solubilidad , Tensoactivos/químicaRESUMEN
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.
Asunto(s)
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
Sulfated polysaccharides (SP) isolated from freshwater green algae, Spirogyra neglecta (Hassall) Kützing, and fractionated SPs were examined to investigate their molecular characteristics and immunomodulatory activity. The crude and fractionated SPs (F1, F2, and F3) consisted mostly of carbohydrates (68.5-85.3%), uronic acids (3.2-4.9%), and sulfates (2.2-12.2%) with various amounts of proteins (2.6-17.1%). D-galactose (23.5-27.3%), D-glucose (11.5-24.8%), L-fucose (19.0-26.7%), and L-rhamnose (16.4-18.3%) were the major monosaccharide units of these SPs with different levels of L-arabinose (3.0-9.4%), D-xylose (4.6-9.8%), and D-mannose (0.4-2.3%). The SPs contained two sub-fractions with molecular weights (Mw) ranging from 164 × 10(3) to 1460 × 10(3) g/mol. The crude and fractionated SPs strongly stimulated murine macrophages, producing considerable amounts of nitric oxide and various cytokines via up-regulation of their mRNA expression by activation of nuclear factor-kappa B and mitogen-activated protein kinases pathways. The main backbone of the most immunoenhancing SP was (1â3)-L-Fucopyranoside, (1â4,6)-D-Glucopyranoside, and (1â4)-D-Galactopyranoside.
Asunto(s)
Citocinas/agonistas , Factores Inmunológicos/química , Macrófagos/efectos de los fármacos , Polisacáridos/química , Spirogyra/química , Proteínas Algáceas/química , Proteínas Algáceas/aislamiento & purificación , Animales , Arabinosa/química , Línea Celular , Citocinas/genética , Citocinas/inmunología , Fucosa/química , Galactosa/química , Regulación de la Expresión Génica , Glucosa/química , Glucósidos/química , Factores Inmunológicos/aislamiento & purificación , Factores Inmunológicos/farmacología , Macrófagos/citología , Macrófagos/inmunología , Manosa/química , Ratones , Proteínas Quinasas Activadas por Mitógenos/genética , Proteínas Quinasas Activadas por Mitógenos/inmunología , FN-kappa B/agonistas , FN-kappa B/genética , FN-kappa B/inmunología , Óxido Nítrico/biosíntesis , Óxido Nítrico/inmunología , Extractos Vegetales/química , Polisacáridos/aislamiento & purificación , Polisacáridos/farmacología , Ramnosa/química , Sulfatos/química , Ácidos Urónicos/química , Ácidos Urónicos/aislamiento & purificación , Xilosa/químicaRESUMEN
Selenium-containing compounds and selenized yeast have anticancer properties. In order to address possible mechanisms involved in these effects, selenoglycoproteins (SGPs) were extracted from selenium-enriched yeast at pH 4.0 and 6.5 (the fractions are called SGP40 and SGP65, respectively), followed by evaluation of their impact on the interactions of lung and breast tumor cells with human brain microvascular endothelial cells (HBMECs). Extracted SGPs, especially SGP40, significantly inhibited adhesion of tumor cells to HBMECs and their transendothelial migration. Because the active components of SGPs are unknown, small selenium-containing compounds [leucyl-valyl-selenomethionyl-arginine (LVSe-MR) and methylselenoadenosine (M-Se-A)], which are normally present in selenized yeast, were introduced as additional treatment groups. Treatment of HBMECs with SGP40, LVSe-MR and M-Se-A induced changes in gene signatures, which suggested a central involvement of nuclear factor (NF)-κB-dependent pathway. These observations were confirmed in the subsequent analysis of NF-κB DNA binding activity, quantitative measurements of the expression of selected genes and proteins, and tumor cell adhesion assay with a specific NF-κB inhibitor as the additional treatment factor. These findings indicate that specific organic selenium-containing compounds have the ability to inhibit tumor cell adhesion to brain endothelial cells via down-regulation of NF-κB. SGPs appear to be more effective than small selenium-containing compounds, suggesting the role of not only selenium but also the glycoprotein component in the observed protective impact.
Asunto(s)
Antineoplásicos/farmacología , Neoplasias de la Mama/tratamiento farmacológico , Endotelio Vascular/efectos de los fármacos , Glicoproteínas/farmacología , Neoplasias Pulmonares/tratamiento farmacológico , Proteínas de Saccharomyces cerevisiae/farmacología , Selenoproteínas/farmacología , Antineoplásicos/aislamiento & purificación , Antineoplásicos/metabolismo , Encéfalo/irrigación sanguínea , Encéfalo/citología , Encéfalo/efectos de los fármacos , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Adhesión Celular/efectos de los fármacos , Línea Celular , Línea Celular Tumoral , Endotelio Vascular/citología , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Glicoproteínas/biosíntesis , Glicoproteínas/aislamiento & purificación , Humanos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Masculino , Microvasos/citología , Microvasos/efectos de los fármacos , FN-kappa B/agonistas , FN-kappa B/genética , FN-kappa B/metabolismo , Proteínas de Neoplasias/agonistas , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Compuestos de Organoselenio/aislamiento & purificación , Compuestos de Organoselenio/metabolismo , Compuestos de Organoselenio/farmacología , Saccharomyces cerevisiae/crecimiento & desarrollo , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/biosíntesis , Proteínas de Saccharomyces cerevisiae/aislamiento & purificación , Selenio/metabolismo , Selenometionina/análogos & derivados , Selenometionina/aislamiento & purificación , Selenometionina/metabolismo , Selenometionina/farmacología , Selenoproteínas/biosíntesis , Selenoproteínas/aislamiento & purificación , Migración Transendotelial y Transepitelial/efectos de los fármacosRESUMEN
The ligation of interleukin-1 receptor (IL-1R) or tumor necrosis factor receptor 1 (TNFR1) induces the recruitment of adaptor proteins and their concomitant ubiquitination to the proximal receptor signaling complex, respectively. Such are upstream signaling events of IKK that play essential roles in NF-κB activation. Thus, the discovery of a substance that would modulate the recruitment of key proximal signaling elements at the upstream level of IKK has been impending in this field of study. Here, we propose that brazilin, an active compound of Caesalpinia sappan L. (Leguminosae), is a potent NF-κB inhibitor that selectively disrupts the formation of the upstream IL-1R signaling complex. Analysis of upstream signaling events revealed that brazilin markedly abolished the IL-1ß-induced polyubiquitination of IRAK1 and its interaction with IKK-γ counterpart. Notably, pretreatment of brazilin drastically interfered the recruitment of the receptor-proximal signaling components including IRAK1/4 and TRAF6 onto MyD88 in IL-1R-triggerd NF-κB activation. Interestingly, brazilin did not affect the TNF-induced RIP1 ubiquitination and the recruitment of RIP1 and TRAF2 to TNFR1, suggesting that brazilin is effective in selectively suppressing the proximal signaling complex formation of IL-1R, but not that of TNFR1. Moreover, our findings suggest that such a disruption of IL-1R-proximal complex formation by brazilin is not mediated by affecting the heterodimerization of IL-1R and IL-1RAcP. Taken together, the results suggest that the anti-IKK activity of brazilin is induced by targeting IKK upstream signaling components and subsequently disrupting proximal IL-1 receptor signaling complex formation.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Benzopiranos/farmacología , Quinasa I-kappa B/antagonistas & inhibidores , Quinasas Asociadas a Receptores de Interleucina-1/antagonistas & inhibidores , Transducción de Señal/efectos de los fármacos , Receptor Toll-Like 4/antagonistas & inhibidores , Antiinflamatorios no Esteroideos/química , Antiinflamatorios no Esteroideos/aislamiento & purificación , Benzopiranos/química , Benzopiranos/aislamiento & purificación , Caesalpinia/química , Etnofarmacología , Genes Reporteros/efectos de los fármacos , Células HEK293 , Células HeLa , Humanos , Quinasa I-kappa B/metabolismo , Proteínas I-kappa B/antagonistas & inhibidores , Proteínas I-kappa B/genética , Proteínas I-kappa B/metabolismo , Quinasas Asociadas a Receptores de Interleucina-1/genética , Quinasas Asociadas a Receptores de Interleucina-1/metabolismo , Interleucina-1beta/antagonistas & inhibidores , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Estructura Molecular , FN-kappa B/agonistas , FN-kappa B/antagonistas & inhibidores , FN-kappa B/genética , FN-kappa B/metabolismo , Receptores de Interleucina-1/agonistas , Receptores de Interleucina-1/antagonistas & inhibidores , Receptores de Interleucina-1/genética , Receptores de Interleucina-1/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , República de Corea , Receptor Toll-Like 4/genética , Receptor Toll-Like 4/metabolismo , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismo , Ubiquitinación/efectos de los fármacos , Madera/químicaRESUMEN
Perilla frutescens has been used in traditional medicine for respiratory diseases due to its anti-bacterial and anti-inflammatory activity. This study aimed to investigate effects of Perilla frutescens leaf extract (PFE) on expression of pro-allergic and pro-inflammatory cytokines in airway epithelial cells exposed to mite major allergen Der p 2 (DP2) and the underlying mechanisms. Our results showed that PFE up to 100 µg/mL had no cytotoxic effect on human bronchial epithelial cell BEAS-2B. Further investigations revealed that PFE dose-dependently diminished mRNA expression of pro-allergic cytokine IL-4, IL-5, IL-13 and GM-CSF, as well as pro-inflammatory cytokine IL-6, IL-8 and MCP-1 in BEAS-2B cells treated with DP2. In parallel to mRNA, the DP-2-elevated levels of the tested cytokines were decreased. Further investigation showed that DP2-indued phosphorylation of p38 MAPK (P38) and JNK, but not Erk1/2, was also suppressed by PFE. In addition, PFE elevated cytosolic IκBα level and decreased nuclear NF-κB level in DP2-stimulated BEAS-2B cells. Taken together, these findings revealed that PFE significantly diminished both mRNA expression and protein levels of pro-allergic and pro-inflammatory cytokines in response to DP2 through inhibition of P38/JNK and NK-κB activation. These findings suggest that PFE should be beneficial to alleviate both allergic and inflammatory responses on airway epithelium in response to aeroallergens.
Asunto(s)
Antígenos Dermatofagoides/farmacología , Proteínas de Artrópodos/farmacología , Citocinas/antagonistas & inhibidores , Células Epiteliales/efectos de los fármacos , Expresión Génica/efectos de los fármacos , Perilla frutescens/química , Extractos Vegetales/farmacología , Hojas de la Planta/química , Animales , Antígenos Dermatofagoides/inmunología , Proteínas de Artrópodos/inmunología , Bronquios/citología , Bronquios/efectos de los fármacos , Bronquios/inmunología , Línea Celular , Citocinas/biosíntesis , Relación Dosis-Respuesta a Droga , Células Epiteliales/citología , Células Epiteliales/inmunología , Humanos , Quinasa I-kappa B/genética , Quinasa I-kappa B/inmunología , MAP Quinasa Quinasa 4/antagonistas & inhibidores , MAP Quinasa Quinasa 4/genética , MAP Quinasa Quinasa 4/inmunología , Ácaros/química , Proteína Quinasa 1 Activada por Mitógenos/genética , Proteína Quinasa 1 Activada por Mitógenos/inmunología , Proteína Quinasa 3 Activada por Mitógenos/genética , Proteína Quinasa 3 Activada por Mitógenos/inmunología , FN-kappa B/agonistas , FN-kappa B/genética , FN-kappa B/inmunología , Extractos Vegetales/aislamiento & purificación , Transducción de Señal , Proteínas Quinasas p38 Activadas por Mitógenos/antagonistas & inhibidores , Proteínas Quinasas p38 Activadas por Mitógenos/genética , Proteínas Quinasas p38 Activadas por Mitógenos/inmunologíaRESUMEN
(R)-(+)-Goniothalamin (GTN), a styryl-lactone isolated from the medicinal plant Goniothalamus macrophyllus, exhibits pharmacological activities including cytotoxic and anti-inflammatory effects. In this study, GTN modulated TNF-α induced NF-κB activation. GTN concentrations up to 20 µM showed low cytotoxic effects in K562 chronic myelogenous leukemia and in Jurkat T cells. Importantly, at these concentrations, no cytotoxicity was observed in healthy peripheral blood mononuclear cells. Our results confirmed that GTN inhibited tumor necrosis factor-α (TNF-α)-induced NF-κB activation in Jurkat and K562 leukemia cells at concentrations as low as 5 µM as shown by reporter gene assays and western blots. Moreover, GTN down-regulated translocation of the p50/p65 heterodimer to the nucleus, prevented binding of NF-κB to its DNA response element and reduced TNF-α-activated interleukin-8 (IL-8) expression. In conclusion, GTN inhibits TNF-α-induced NF-κB activation at non-apoptogenic concentrations in different leukemia cell models without presenting toxicity towards healthy blood cells underlining the anti-leukemic potential of this natural compound.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Antineoplásicos Fitogénicos/farmacología , Descubrimiento de Drogas , Leucemia/tratamiento farmacológico , FN-kappa B/metabolismo , Pironas/farmacología , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Antiinflamatorios no Esteroideos/efectos adversos , Antiinflamatorios no Esteroideos/aislamiento & purificación , Antineoplásicos Fitogénicos/efectos adversos , Antineoplásicos Fitogénicos/aislamiento & purificación , Núcleo Celular/efectos de los fármacos , Núcleo Celular/metabolismo , Células Cultivadas , Genes Reporteros/efectos de los fármacos , Goniothalamus/química , Humanos , Interleucina-8/antagonistas & inhibidores , Interleucina-8/metabolismo , Células Jurkat , Células K562 , Leucocitos Mononucleares/citología , Leucocitos Mononucleares/efectos de los fármacos , Malasia , FN-kappa B/agonistas , FN-kappa B/antagonistas & inhibidores , Proteínas de Neoplasias/antagonistas & inhibidores , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Raíces de Plantas/química , Transporte de Proteínas/efectos de los fármacos , Pironas/efectos adversos , Pironas/aislamiento & purificación , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Elementos de Respuesta/efectos de los fármacos , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Extracts of four plant portions (roots, stems, leaves and flowers) of Urtica dioica (the stinging nettle) were prepared using accelerated solvent extraction (ASE) involving water, hexanes, methanol and dichloromethane. The extracts were evaluated for their anti-inflammatory and cytotoxic activities in an NF-κB luciferase and MTT assay using macrophage immune (RAW264.7) cells. A standardized commercial ethanol extract of nettle leaves was also evaluated. The methanolic extract of the flowering portions displayed significant anti-inflammatory activity on par with a standard compound celastrol (1) but were moderately cytotoxic. Alternatively, the polar extracts (water, methanol, ethanol) of the roots, stems and leaves displayed moderate to weak anti-inflammatory activity, while the methanol and especially the water soluble extracts exhibited noticeable cytotoxicity. In contrast, the lipophilic dichloromethane extracts of the roots, stems and leaves exhibited potent anti-inflammatory effects greater than or equal to 1 with minimal cytotoxicity to RAW264.7 cells. Collectively these results suggest that using lipophilic extracts of stinging nettle may be more effective than traditional tinctures (water, methanol, ethanol) in clinical evaluations for the treatment of inflammatory disorders especially arthritis. A chemical investigation into the lipophilic extracts of stinging nettle to identify the bioactive compound(s) responsible for their observed anti-inflammatory activity is further warranted.
Asunto(s)
Antiinflamatorios/farmacología , Citotoxinas/farmacología , Macrófagos/efectos de los fármacos , Extractos Vegetales/farmacología , Urtica dioica/química , Animales , Antiinflamatorios/química , Citotoxinas/química , Flores/química , Luciferasas/metabolismo , Macrófagos/inmunología , Ratones , Estructura Molecular , FN-kappa B/agonistas , Extractos Vegetales/química , Hojas de la Planta/química , Raíces de Plantas/química , Tallos de la Planta/químicaRESUMEN
UNLABELLED: Liver progenitor cells (LPCs) represent the cell compartment facilitating hepatic regeneration during chronic injury while hepatocyte-mediated repair mechanisms are compromised. LPC proliferation is frequently observed in human chronic liver diseases such as hereditary hemochromatosis, fatty liver disease, and chronic hepatitis. In vivo studies have suggested that a tumor necrosis factor family member, tumor necrosis factor-like weak inducer of apoptosis (TWEAK), is promitotic for LPCs; whether it acts directly is not known. In our murine choline-deficient, ethionine-supplemented (CDE) model of chronic liver injury, TWEAK receptor [fibroblast growth factor-inducible 14 (Fn14)] expression in the whole liver is massively upregulated. We therefore set out to investigate whether TWEAK/Fn14 signaling promotes the regenerative response in CDE-induced chronic liver injury by mitotic stimulation of LPCs. Fn14 knockout (KO) mice showed significantly reduced LPC numbers and attenuated inflammation and cytokine production after 2 weeks of CDE feeding. The close association between LPC proliferation and activation of hepatic stellate cells in chronic liver injury prompted us to investigate whether fibrogenesis was also modulated in Fn14 KO animals. Collagen deposition and expression of key fibrogenesis mediators were reduced after 2 weeks of injury, and this correlated with LPC numbers. Furthermore, the injection of 2-week-CDE-treated wildtype animals with TWEAK led to increased proliferation of nonparenchymal pan cytokeratin-positive cells. Stimulation of an Fn14-positive LPC line with TWEAK led to nuclear factor kappa light chain enhancer of activated B cells (NFkappaB) activation and dose-dependent proliferation, which was diminished after targeting of the p50 NFkappaB subunit by RNA interference. CONCLUSION: TWEAK acts directly and stimulates LPC mitosis in an Fn14-dependent and NFkappaB-dependent fashion, and signaling via this pathway mediates the LPC response to CDE-induced injury and regeneration.
Asunto(s)
Regeneración Hepática , Mitosis , Células Madre/fisiología , Factores de Necrosis Tumoral/fisiología , Animales , Proliferación Celular , Deficiencia de Colina/metabolismo , Colágeno/metabolismo , Citocina TWEAK , Etionina/farmacología , Hígado/efectos de los fármacos , Hígado/lesiones , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Mitógenos/farmacología , FN-kappa B/agonistas , FN-kappa B/genética , FN-kappa B/metabolismo , Receptores del Factor de Necrosis Tumoral/genética , Proteínas Recombinantes/farmacología , Células Madre/efectos de los fármacos , Receptor de TWEAK , Factores de Necrosis Tumoral/farmacologíaRESUMEN
INTRODUCTION: The ability to predict the health effects resulting from drug or chemical exposure has been challenging due to the complexity of human biology. Approaches that detect and discriminate a broad range of mechanisms in testing formats that are predictive and yet cost-effective are needed. METHODS: Here, we evaluated the performance of BioMAP systems, primary human cell-based disease models, as a platform for characterization of chemical toxicity mechanisms. For this we tested a set of compounds with known or well-studied mechanisms in a panel of 8 BioMAP assays relevant to human respiratory, skin, immune and vascular exposure sites. RESULTS: We evaluated the ability to detect and distinguish compounds based on mechanisms of action, comparing the BioMAP activity profiles generated in a reduced sample number format to reference database profiles derived from multiple experiments. We also studied the role of BioMAP assay panel size and concentration effects, both of which were found to contribute to the ability to discriminate chemicals and mechanisms. DISCUSSION: Compounds with diverse mechanisms, including modulators of the NFkappaB pathway, microtubule function and mitochondrial activity, could be discriminated and classified into target and pathway mechanisms in both assay formats. Certain inhibitors of mitochondrial function, such as rotenone and sodium azide, but not others, were classified with inducers of endoplasmic reticulum stress, providing insight into the toxicity mechanisms of these agents. This method may have utility in classifying novel agents with unknown modes of action according to their effects on toxicity pathways.
Asunto(s)
Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos , Exposición a Riesgos Ambientales/efectos adversos , Noxas/clasificación , Preparaciones Farmacéuticas/clasificación , Pruebas de Toxicidad , Biomarcadores , Técnicas de Cultivo de Célula/economía , Células Cultivadas , Evaluación Preclínica de Medicamentos/métodos , Retículo Endoplásmico/efectos de los fármacos , Humanos , Microtúbulos/efectos de los fármacos , Mitocondrias/efectos de los fármacos , FN-kappa B/agonistas , FN-kappa B/antagonistas & inhibidoresRESUMEN
Epilobium angustifolium has been traditionally used to treat of a number of diseases; however, not much is known regarding its effect on innate immune cells. In this study, we report that extracts of E. angustifolium activated functional responses in neutrophils and monocyte/macrophages. Activity-guided fractionation, followed by mass spectroscopy and NMR analysis, resulted in the identification of oenothein B as the primary component responsible for phagocyte activation. Oenothein B, a dimeric hydrolysable tannin, dose-dependently induced a number of phagocyte functions in vitro, including intracellular Ca(2+) flux, production of reactive oxygen species, chemotaxis, NF-kappaB activation, and proinflammatory cytokine production. Furthermore, oenothein B was active in vivo, inducing keratinocyte chemoattractant production and neutrophil recruitment to the peritoneum after intraperitoneal administration. Biological activity required the full oenothein B structure, as substructures of oenothein B (pyrocatechol, gallic acid, pyrogallol, 3,4-dihydroxybenzoic acid) were all inactive. The ability of oenothein B to modulate phagocyte functions in vitro and in vivo suggests that this compound is responsible for at least part of the therapeutic properties of E. angustifolium extracts.
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Taninos Hidrolizables/farmacología , Factores Inmunológicos/farmacología , Activación de Macrófagos , Macrófagos/efectos de los fármacos , FN-kappa B/inmunología , Animales , Calcio/agonistas , Calcio/inmunología , Calcio/metabolismo , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Movimiento Celular/inmunología , Quimiotaxis/efectos de los fármacos , Quimiotaxis/inmunología , Citocinas/efectos de los fármacos , Citocinas/inmunología , Citocinas/metabolismo , Epilobium/química , Femenino , Humanos , Taninos Hidrolizables/aislamiento & purificación , Factores Inmunológicos/aislamiento & purificación , Macrófagos/inmunología , Ratones , Ratones Endogámicos BALB C , FN-kappa B/agonistas , FN-kappa B/metabolismo , Neutrófilos/efectos de los fármacos , Neutrófilos/inmunología , Neutrófilos/metabolismo , Extractos Vegetales/química , Extractos Vegetales/farmacología , Especies Reactivas de Oxígeno/agonistas , Especies Reactivas de Oxígeno/inmunología , Especies Reactivas de Oxígeno/metabolismoAsunto(s)
Antiinflamatorios no Esteroideos/uso terapéutico , Dermatitis Atópica/tratamiento farmacológico , Lithospermum/química , Extractos Vegetales/uso terapéutico , Administración Oral , Animales , Enfermedad Crónica , Ciclooxigenasa 2/efectos de los fármacos , Ciclooxigenasa 2/metabolismo , Dermatitis Atópica/inducido químicamente , Femenino , Proteínas I-kappa B/efectos de los fármacos , Proteínas I-kappa B/metabolismo , Ratones , Inhibidor NF-kappaB alfa , FN-kappa B/agonistas , FN-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo II/efectos de los fármacos , Óxido Nítrico Sintasa de Tipo II/metabolismo , Oxazolona/farmacología , Fitoterapia , Prednisolona/uso terapéutico , Piel/efectos de los fármacos , Piel/patologíaRESUMEN
Clerodendron trichotomum Tunberg Leaves (CTL) have been used for centuries in Chinese folk medicine for their anti-inflammatory properties. To investigate the molecular mechanism of anti-inflammation by CTL, we analyzed the regulation of TNF-alpha expression in RAW 264.7 cells, a key step in inflammation. The effect of CTL on the production and expression of tumor necrosis factor-alpha (TNF-alpha) was determined by enzyme-linked immunosorbent assay (ELISA) and reverse transcription polymerase chain reaction (RT-PCR). CTL inhibited the production and expression of TNF-alpha in LPS-stimulated RAW 264.7 cells in a dose-dependent manner. In addition, activation of NF-kappaB, which controls TNF-alpha expression, was inhibited in LPS-stimulated RAW 264.7 cells by CTL in a dose-dependent manner, as demonstrated by an electro phoretic mobility shift assay (EMSA). Furthermore, CTL inhibited activation of NF-kappaB through inhibition IkappaB degradation, as demonstrated by an western blot analysis of IkappaB-alpha. These results suggest that CTL inhibits the expression of the pro-inflammation gene through the inhibition of NF-kappaB dependent pathway in RAW 264.7 cells.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Clerodendrum/química , Inflamación/genética , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , FN-kappa B/agonistas , Animales , Antiinflamatorios no Esteroideos/aislamiento & purificación , Western Blotting , Línea Celular , Ensayo de Cambio de Movilidad Electroforética , Ensayo de Inmunoadsorción Enzimática , Ratones , Hojas de la Planta/química , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Sales de Tetrazolio , Tiazoles , Factor de Necrosis Tumoral alfa/biosíntesisRESUMEN
Chelidonium majus is used to treat several inflammatory diseases and tumours. We have examined the effect of C. majus on nitric oxide (NO) production using mouse peritoneal macrophages. When C. majus was used in combination with recombinant interferon-gamma (rIFN-gamma, 10 U mL(-1)), there was a marked cooperative induction of NO production. Treatment of rIFN-gamma plus C. majus (1 mgmL(-1)) in macrophages caused a significant increase in tumour necrosis factor-alpha (TNF-alpha) production. The increased production of NO and TNF-alpha from rIFN-gamma plus C. majus-stimulated cells was almost completely inhibited by nuclear factor-kappaB (NF-kappaB) inhibitor, pyrrolidine dithiocarbamate (100 microM). These findings demonstrated that C. majus increased the production of NO and TNF-alpha by rIFN-gamma-primed macrophages and suggested that NF-kappaB played a critical role in mediating the effects of C. majus.