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1.
Food Funct ; 9(9): 4865-4875, 2018 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-30160278

RESUMO

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.


Assuntos
Conservadores da Densidade Óssea/uso terapêutico , Condrócitos/metabolismo , Suplementos Nutricionais , Modelos Animais de Doenças , Osteoartrite do Joelho/terapia , Inibidores de Fosfoinositídeo-3 Quinase , Ácidos Sulfínicos/uso terapêutico , Animais , Conservadores da Densidade Óssea/efeitos adversos , Conservadores da Densidade Óssea/metabolismo , Sobrevivência Celular , Células Cultivadas , Condrócitos/citologia , Condrócitos/patologia , Suplementos Nutricionais/efeitos adversos , Progressão da Doença , Dissulfetos , Feminino , Regulação da Expressão Gênica , Humanos , Interleucina-1beta/antagonistas & inibidores , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Masculino , Camundongos Endogâmicos C57BL , NF-kappa B/agonistas , NF-kappa B/antagonistas & inibidores , NF-kappa B/metabolismo , Osteoartrite do Joelho/metabolismo , Osteoartrite do Joelho/patologia , Osteoartrite do Joelho/fisiopatologia , Fosfatidilinositol 3-Quinase/química , Fosfatidilinositol 3-Quinase/metabolismo , Proteínas Proto-Oncogênicas c-akt/agonistas , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-akt/metabolismo , Distribuição Aleatória , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Transdução de Sinais , Ácidos Sulfínicos/efeitos adversos , Ácidos Sulfínicos/metabolismo
2.
Sci Rep ; 6: 27566, 2016 06 14.
Artigo em Inglês | MEDLINE | ID: mdl-27297123

RESUMO

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.


Assuntos
Antituberculosos/farmacologia , Dicumarol/farmacologia , Interações Hospedeiro-Patógeno/efeitos dos fármacos , Macrófagos/imunologia , Mycobacterium tuberculosis/efeitos dos fármacos , NAD(P)H Desidrogenase (Quinona)/genética , Sinergismo Farmacológico , Inibidores Enzimáticos/farmacologia , Regulação da Expressão Gênica , Biblioteca Gênica , Sequenciamento de Nucleotídeos em Larga Escala , Ensaios de Triagem em Larga Escala , Humanos , Interleucina-1beta/agonistas , Interleucina-1beta/genética , Interleucina-1beta/imunologia , Macrófagos/efeitos dos fármacos , Macrófagos/microbiologia , Mycobacterium tuberculosis/patogenicidade , Mycobacterium tuberculosis/fisiologia , NAD(P)H Desidrogenase (Quinona)/antagonistas & inibidores , NAD(P)H Desidrogenase (Quinona)/imunologia , NF-kappa B/agonistas , NF-kappa B/genética , NF-kappa B/imunologia , Cultura Primária de Células , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Rifampina/farmacologia , Transdução de Sinais , Células THP-1 , Fator de Necrose Tumoral alfa/agonistas , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia
3.
Basic Clin Pharmacol Toxicol ; 119(6): 540-547, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27194111

RESUMO

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.


Assuntos
Antimetabólitos Antineoplásicos/efeitos adversos , Antioxidantes/uso terapêutico , Colo Descendente/efeitos dos fármacos , Curcumina/uso terapêutico , Diarreia/prevenção & controle , Suplementos Nutricionais , Fluoruracila/efeitos adversos , Animais , Anti-Inflamatórios não Esteroides/administração & dosagem , Anti-Inflamatórios não Esteroides/farmacologia , Anti-Inflamatórios não Esteroides/uso terapêutico , Antimetabólitos Antineoplásicos/química , Antimetabólitos Antineoplásicos/farmacologia , Antioxidantes/administração & dosagem , Antioxidantes/farmacologia , Quimiocina CXCL1/agonistas , Quimiocina CXCL1/antagonistas & inibidores , Quimiocina CXCL1/genética , Quimiocina CXCL1/metabolismo , Quimiocina CXCL2/agonistas , Quimiocina CXCL2/antagonistas & inibidores , Quimiocina CXCL2/genética , Quimiocina CXCL2/metabolismo , Colo Descendente/imunologia , Colo Descendente/metabolismo , Colo Descendente/fisiopatologia , Curcumina/administração & dosagem , Curcumina/farmacologia , Diarreia/induzido quimicamente , Diarreia/metabolismo , Diarreia/fisiopatologia , Proteína p300 Associada a E1A/agonistas , Proteína p300 Associada a E1A/antagonistas & inibidores , Proteína p300 Associada a E1A/metabolismo , Inibidores Enzimáticos/farmacologia , Fluoruracila/antagonistas & inibidores , Fluoruracila/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/imunologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/fisiopatologia , Masculino , Camundongos Endogâmicos C57BL , NF-kappa B/agonistas , NF-kappa B/antagonistas & inibidores , NF-kappa B/metabolismo , Índice de Gravidade de Doença , Técnicas de Cultura de Tecidos
4.
Biosci Biotechnol Biochem ; 80(7): 1393-402, 2016 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-27095137

RESUMO

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.


Assuntos
Adjuvantes Imunológicos/farmacologia , Interleucina-6/agonistas , Macrófagos Peritoneais/efeitos dos fármacos , Extratos Vegetais/farmacologia , Spinacia oleracea/química , Fator de Necrose Tumoral alfa/agonistas , Animais , Linhagem Celular , Feminino , Expressão Gênica , Interleucina-6/genética , Interleucina-6/imunologia , Macrófagos Peritoneais/citologia , Macrófagos Peritoneais/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/imunologia , NF-kappa B/agonistas , NF-kappa B/genética , NF-kappa B/imunologia , Fagocitose/efeitos dos fármacos , Cultura Primária de Células , RNA Mensageiro/agonistas , RNA Mensageiro/genética , RNA Mensageiro/imunologia , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/imunologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia
5.
Food Chem Toxicol ; 85: 2-9, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26260749

RESUMO

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.


Assuntos
Anti-Inflamatórios não Esteroides/metabolismo , Enterócitos/metabolismo , Absorção Intestinal , NF-kappa B/metabolismo , Nanopartículas/efeitos adversos , Triterpenos Pentacíclicos/metabolismo , Poliglactina 910/efeitos adversos , Transporte Ativo do Núcleo Celular , Algoritmos , Anti-Inflamatórios não Esteroides/administração & dosagem , Anti-Inflamatórios não Esteroides/análise , Anti-Inflamatórios não Esteroides/química , Células CACO-2 , Permeabilidade da Membrana Celular , Sobrevivência Celular , Fenômenos Químicos , Suplementos Nutricionais/análise , Emulsões , Humanos , Microscopia Eletrônica de Transmissão , NF-kappa B/agonistas , NF-kappa B/antagonistas & inibidores , Nanopartículas/química , Nanopartículas/ultraestrutura , Tamanho da Partícula , Triterpenos Pentacíclicos/administração & dosagem , Triterpenos Pentacíclicos/análise , Triterpenos Pentacíclicos/química , Poliglactina 910/química , Polivinil/química , Solubilidade , Tensoativos/química
6.
Mol Med Rep ; 12(4): 5580-6, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26166377

RESUMO

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.


Assuntos
Proteínas Quinases Ativadas por AMP/genética , Anti-Inflamatórios não Esteroides/farmacologia , Berberina/farmacologia , Células Endoteliais/efeitos dos fármacos , NF-kappa B/genética , Fator de Necrose Tumoral alfa/farmacologia , Proteínas Quinases Ativadas por AMP/antagonistas & inibidores , Proteínas Quinases Ativadas por AMP/metabolismo , Linhagem Celular , Quimiocina CCL2/agonistas , Quimiocina CCL2/antagonistas & inibidores , Quimiocina CCL2/genética , Quimiocina CCL2/metabolismo , Células Endoteliais/citologia , Células Endoteliais/metabolismo , Regulação da Expressão Gênica , Humanos , Inflamação , Molécula 1 de Adesão Intercelular/genética , Molécula 1 de Adesão Intercelular/metabolismo , NF-kappa B/agonistas , NF-kappa B/antagonistas & inibidores , NF-kappa B/metabolismo , Inibidores de Proteínas Quinases/farmacologia , RNA Mensageiro/agonistas , RNA Mensageiro/antagonistas & inibidores , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Transdução de Sinais , Fator de Necrose Tumoral alfa/antagonistas & inibidores
7.
Biosci Biotechnol Biochem ; 79(10): 1644-53, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25971153

RESUMO

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 (1→3)-L-Fucopyranoside, (1→4,6)-D-Glucopyranoside, and (1→4)-D-Galactopyranoside.


Assuntos
Citocinas/agonistas , Fatores Imunológicos/química , Macrófagos/efeitos dos fármacos , Polissacarídeos/química , Spirogyra/química , Proteínas de Algas/química , Proteínas de Algas/isolamento & purificação , Animais , Arabinose/química , Linhagem Celular , Citocinas/genética , Citocinas/imunologia , Fucose/química , Galactose/química , Regulação da Expressão Gênica , Glucose/química , Glucosídeos/química , Fatores Imunológicos/isolamento & purificação , Fatores Imunológicos/farmacologia , Macrófagos/citologia , Macrófagos/imunologia , Manose/química , Camundongos , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/imunologia , NF-kappa B/agonistas , NF-kappa B/genética , NF-kappa B/imunologia , Óxido Nítrico/biossíntese , Óxido Nítrico/imunologia , Extratos Vegetais/química , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologia , Ramnose/química , Sulfatos/química , Ácidos Urônicos/química , Ácidos Urônicos/isolamento & purificação , Xilose/química
8.
J Nutr Biochem ; 26(2): 120-9, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25465156

RESUMO

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.


Assuntos
Antineoplásicos/farmacologia , Neoplasias da Mama/tratamento farmacológico , Endotélio Vascular/efeitos dos fármacos , Glicoproteínas/farmacologia , Neoplasias Pulmonares/tratamento farmacológico , Proteínas de Saccharomyces cerevisiae/farmacologia , Selenoproteínas/farmacologia , Antineoplásicos/isolamento & purificação , Antineoplásicos/metabolismo , Encéfalo/irrigação sanguínea , Encéfalo/citologia , Encéfalo/efeitos dos fármacos , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Adesão Celular/efeitos dos fármacos , Linhagem Celular , Linhagem Celular Tumoral , Endotélio Vascular/citologia , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Glicoproteínas/biossíntese , Glicoproteínas/isolamento & purificação , Humanos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Masculino , Microvasos/citologia , Microvasos/efeitos dos fármacos , NF-kappa B/agonistas , NF-kappa B/genética , NF-kappa B/metabolismo , Proteínas de Neoplasias/agonistas , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Compostos Organosselênicos/isolamento & purificação , Compostos Organosselênicos/metabolismo , Compostos Organosselênicos/farmacologia , Saccharomyces cerevisiae/crescimento & desenvolvimento , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/biossíntese , Proteínas de Saccharomyces cerevisiae/isolamento & purificação , Selênio/metabolismo , Selenometionina/análogos & derivados , Selenometionina/isolamento & purificação , Selenometionina/metabolismo , Selenometionina/farmacologia , Selenoproteínas/biossíntese , Selenoproteínas/isolamento & purificação , Migração Transendotelial e Transepitelial/efeitos dos fármacos
9.
Biochem Pharmacol ; 89(4): 515-25, 2014 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-24735611

RESUMO

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.


Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Benzopiranos/farmacologia , Quinase I-kappa B/antagonistas & inibidores , Quinases Associadas a Receptores de Interleucina-1/antagonistas & inibidores , Transdução de Sinais/efeitos dos fármacos , Receptor 4 Toll-Like/antagonistas & inibidores , Anti-Inflamatórios não Esteroides/química , Anti-Inflamatórios não Esteroides/isolamento & purificação , Benzopiranos/química , Benzopiranos/isolamento & purificação , Caesalpinia/química , Etnofarmacologia , Genes Reporter/efeitos dos fármacos , Células HEK293 , Células HeLa , Humanos , Quinase I-kappa B/metabolismo , Proteínas I-kappa B/antagonistas & inibidores , Proteínas I-kappa B/genética , Proteínas I-kappa B/metabolismo , Quinases Associadas a Receptores de Interleucina-1/genética , Quinases Associadas a Receptores de Interleucina-1/metabolismo , Interleucina-1beta/antagonistas & inibidores , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Estrutura Molecular , NF-kappa B/agonistas , NF-kappa B/antagonistas & inibidores , NF-kappa B/genética , NF-kappa B/metabolismo , Receptores de Interleucina-1/agonistas , Receptores de Interleucina-1/antagonistas & inibidores , Receptores de Interleucina-1/genética , Receptores de Interleucina-1/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , República da Coreia , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/metabolismo , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Ubiquitinação/efeitos dos fármacos , Madeira/química
10.
PLoS One ; 8(10): e77458, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24204835

RESUMO

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.


Assuntos
Antígenos de Dermatophagoides/farmacologia , Proteínas de Artrópodes/farmacologia , Citocinas/antagonistas & inibidores , Células Epiteliais/efeitos dos fármacos , Expressão Gênica/efeitos dos fármacos , Perilla frutescens/química , Extratos Vegetais/farmacologia , Folhas de Planta/química , Animais , Antígenos de Dermatophagoides/imunologia , Proteínas de Artrópodes/imunologia , Brônquios/citologia , Brônquios/efeitos dos fármacos , Brônquios/imunologia , Linhagem Celular , Citocinas/biossíntese , Relação Dose-Resposta a Droga , Células Epiteliais/citologia , Células Epiteliais/imunologia , Humanos , Quinase I-kappa B/genética , Quinase I-kappa B/imunologia , MAP Quinase Quinase 4/antagonistas & inibidores , MAP Quinase Quinase 4/genética , MAP Quinase Quinase 4/imunologia , Ácaros/química , Proteína Quinase 1 Ativada por Mitógeno/genética , Proteína Quinase 1 Ativada por Mitógeno/imunologia , Proteína Quinase 3 Ativada por Mitógeno/genética , Proteína Quinase 3 Ativada por Mitógeno/imunologia , NF-kappa B/agonistas , NF-kappa B/genética , NF-kappa B/imunologia , Extratos Vegetais/isolamento & purificação , Transdução de Sinais , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases p38 Ativadas por Mitógeno/genética , Proteínas Quinases p38 Ativadas por Mitógeno/imunologia
11.
Food Chem Toxicol ; 59: 572-8, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23845509

RESUMO

(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.


Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Antineoplásicos Fitogênicos/farmacologia , Descoberta de Drogas , Leucemia/tratamento farmacológico , NF-kappa B/metabolismo , Pironas/farmacologia , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Anti-Inflamatórios não Esteroides/efeitos adversos , Anti-Inflamatórios não Esteroides/isolamento & purificação , Antineoplásicos Fitogênicos/efeitos adversos , Antineoplásicos Fitogênicos/isolamento & purificação , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Células Cultivadas , Genes Reporter/efeitos dos fármacos , Goniothalamus/química , Humanos , Interleucina-8/antagonistas & inibidores , Interleucina-8/metabolismo , Células Jurkat , Células K562 , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/efeitos dos fármacos , Malásia , NF-kappa B/agonistas , NF-kappa B/antagonistas & inibidores , Proteínas de Neoplasias/antagonistas & inibidores , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Raízes de Plantas/química , Transporte Proteico/efeitos dos fármacos , Pironas/efeitos adversos , Pironas/isolamento & purificação , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Elementos de Resposta/efeitos dos fármacos , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo
12.
Phytomedicine ; 20(2): 143-7, 2013 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-23092723

RESUMO

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.


Assuntos
Anti-Inflamatórios/farmacologia , Citotoxinas/farmacologia , Macrófagos/efeitos dos fármacos , Extratos Vegetais/farmacologia , Urtica dioica/química , Animais , Anti-Inflamatórios/química , Citotoxinas/química , Flores/química , Luciferases/metabolismo , Macrófagos/imunologia , Camundongos , Estrutura Molecular , NF-kappa B/agonistas , Extratos Vegetais/química , Folhas de Planta/química , Raízes de Plantas/química , Caules de Planta/química
13.
Hepatology ; 52(1): 291-302, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20578156

RESUMO

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.


Assuntos
Regeneração Hepática , Mitose , Células-Tronco/fisiologia , Fatores de Necrose Tumoral/fisiologia , Animais , Proliferação de Células , Deficiência de Colina/metabolismo , Colágeno/metabolismo , Citocina TWEAK , Etionina/farmacologia , Fígado/efeitos dos fármacos , Fígado/lesões , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Mitógenos/farmacologia , NF-kappa B/agonistas , NF-kappa B/genética , NF-kappa B/metabolismo , Receptores do Fator de Necrose Tumoral/genética , Proteínas Recombinantes/farmacologia , Células-Tronco/efeitos dos fármacos , Receptor de TWEAK , Fatores de Necrose Tumoral/farmacologia
14.
J Pharmacol Toxicol Methods ; 61(1): 3-15, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-19879948

RESUMO

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.


Assuntos
Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Exposição Ambiental/efeitos adversos , Noxas/classificação , Preparações Farmacêuticas/classificação , Testes de Toxicidade , Biomarcadores , Técnicas de Cultura de Células/economia , Células Cultivadas , Avaliação Pré-Clínica de Medicamentos/métodos , Retículo Endoplasmático/efeitos dos fármacos , Humanos , Microtúbulos/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , NF-kappa B/agonistas , NF-kappa B/antagonistas & inibidores
15.
J Immunol ; 183(10): 6754-66, 2009 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-19846877

RESUMO

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.


Assuntos
Taninos Hidrolisáveis/farmacologia , Fatores Imunológicos/farmacologia , Ativação de Macrófagos , Macrófagos/efeitos dos fármacos , NF-kappa B/imunologia , Animais , Cálcio/agonistas , Cálcio/imunologia , Cálcio/metabolismo , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Movimento Celular/imunologia , Quimiotaxia/efeitos dos fármacos , Quimiotaxia/imunologia , Citocinas/efeitos dos fármacos , Citocinas/imunologia , Citocinas/metabolismo , Epilobium/química , Feminino , Humanos , Taninos Hidrolisáveis/isolamento & purificação , Fatores Imunológicos/isolamento & purificação , Macrófagos/imunologia , Camundongos , Camundongos Endogâmicos BALB C , NF-kappa B/agonistas , NF-kappa B/metabolismo , Neutrófilos/efeitos dos fármacos , Neutrófilos/imunologia , Neutrófilos/metabolismo , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Espécies Reativas de Oxigênio/agonistas , Espécies Reativas de Oxigênio/imunologia , Espécies Reativas de Oxigênio/metabolismo
17.
Arch Pharm Res ; 30(6): 755-60, 2007 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-17679555

RESUMO

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.


Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Clerodendrum/química , Inflamação/genética , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , NF-kappa B/agonistas , Animais , Anti-Inflamatórios não Esteroides/isolamento & purificação , Western Blotting , Linhagem Celular , Ensaio de Desvio de Mobilidade Eletroforética , Ensaio de Imunoadsorção Enzimática , Camundongos , Folhas de Planta/química , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sais de Tetrazólio , Tiazóis , Fator de Necrose Tumoral alfa/biossíntese
18.
J Pharm Pharmacol ; 56(1): 129-34, 2004 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-14980010

RESUMO

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.


Assuntos
Chelidonium , NF-kappa B/agonistas , Óxido Nítrico/biossíntese , Extratos Vegetais/farmacologia , Fator de Necrose Tumoral alfa/biossíntese , Animais , Western Blotting , Técnicas In Vitro , Interferon gama/farmacologia , Macrófagos Peritoneais/metabolismo , Camundongos , NF-kappa B/antagonistas & inibidores , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico Sintase/biossíntese , Óxido Nítrico Sintase Tipo II , Extratos Vegetais/metabolismo , Pirrolidinas/farmacologia , Proteínas Recombinantes , Tiocarbamatos/farmacologia , ômega-N-Metilarginina/farmacologia
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