RESUMO
Sepsis is characterized by an initial net hyperinflammatory response, followed by a period of immunosuppression, termed immunoparalysis. During this immunosuppressive phase, patients may have difficulty eradicating invading pathogens and are susceptible to life-threatening secondary hospital-acquired infections. Due to progress in antimicrobial treatment and supportive care, most patients survive early sepsis. Mortality is more frequently attributed to subsequent secondary nosocomial infections and multiorgan system failure. 6-Gingerol is the major pharmacologically active component of ginger. Although it is known to exhibit a variety of biological activities, including anti-inflammation and antioxidation, the role of 6-gingerol in sepsis-induced immune dysfunction remains elusive. Thus, we investigated whether 6-gingerol improves septic host response to infections during sepsis. 6-Gingerol-treated mice showed significantly lower mortality in polymicrobial sepsis induced by cecal ligation and puncture LPS via enhanced bacterial clearance in the peritoneum, blood, and organs (liver, spleen, and kidney) and inhibited the production of TNF-α and IL-6 in TLR2 and/or TLR4-stimulated macrophages. In addition, we demonstrated that survival improvement of secondary infection following septic insult was associated with an initial response of enhanced neutrophil numbers and function at the infection site, reduced apoptosis of immune cells, and a shift from a T helper cell type 2 (Th2) to a T helper cell type 1 (Th1) cytokine balance in the hypoinflammation phase. Our overall findings suggest that 6-gingerol potentially restores sepsis-induced immune dysfunction by shifting the balance of Th1/Th2 and by regulating apoptosis of immune cells.
Assuntos
Catecóis/uso terapêutico , Citocinas/metabolismo , Álcoois Graxos/uso terapêutico , Doenças do Sistema Imunitário/tratamento farmacológico , Linfócitos/metabolismo , Sepse/complicações , Animais , Apoptose , Catecóis/farmacologia , Álcoois Graxos/farmacologia , Humanos , Doenças do Sistema Imunitário/fisiopatologia , Masculino , CamundongosRESUMO
The translocation of transcription factor EB (TFEB) to the nucleus plays a pivotal role in the regulation of basic cellular processes, such as lysosome biogenesis and autophagy. Autophagy is an intracellular degradation system that delivers cytoplasmic constituents to the lysosome, which is important in maintaining cellular homeostasis during environmental stress. Furthermore, oxidative stress is a critical cause for the progression of neurodegenerative diseases. Curcumin has anti-oxidative and anti-inflammatory activities, and is expected to have potential therapeutic effects in various diseases. In this study, we demonstrated that curcumin regulated TFEB export signalling via inhibition of glycogen synthase kinase-3ß (GSK-3ß); GSK-3ß was inactivated by curcumin, leading to reduced phosphorylation of TFEB. We further showed that H2O2-induced oxidative stress was reduced by curcumin via the Nrf2/HO-1 pathway in human neuroblastoma cells. In addition, we showed that curcumin induced the degradation of amyloidogenic proteins, including amyloid-ß precursor protein and α-synuclein, through the TFEB-autophagy/lysosomal pathway. In conclusion, curcumin regulates autophagy by controlling TFEB through the inhibition of GSK-3ß, and increases antioxidant gene expression in human neuroblastoma cells. These results contribute to the development of novel cellular therapies for neurodegenerative diseases.
Assuntos
Precursor de Proteína beta-Amiloide/metabolismo , Antineoplásicos/uso terapêutico , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/metabolismo , Curcumina/uso terapêutico , Glicogênio Sintase Quinase 3 beta/antagonistas & inibidores , Neuroblastoma/genética , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Curcumina/farmacologia , Humanos , Espécies Reativas de Oxigênio , TransfecçãoRESUMO
Chung Hun Wha Dam Tang (CHWDT), a traditional Korean herbal formula, has been used for hundreds of years for alleviating dizziness, phlegm, and inflammation. The inhibitory effects of CHWDT on obesity have been reported. However, the effects of CHWDT in atherosclerosis have not yet been explored. Therefore, the aim of the study was to investigate whether CHWDT could confer protection from oxidative stress and inflammation in a high fat diet (HFD)-induced atherosclerosis model. Atherosclerosis was induced by feeding ApoeE-/- mice with HFD for 6 weeks. To examine the in vivo effects of CHWDT on HFD-induced atherosclerosis, mice on HFD for 6 weeks were orally administrated with CHWDT (400 or 800â¯mg/kg) every other day for an additional 6 weeks and histological features of aorta were determined by Sudan IV and H&E staining. The mRNA levels of TNF-α, SOD1, SOD2, iNOS or eNOS were determined with RT-PCR analysis or western blot analysis for protein levels. ROS generation was measured by CM-2DCFDA or MitoSox staining using FACS analysis or confocal microscopy. CHWDT decreased the mRNA levels of TNF-α and increased the mRNA levels of SOD1, SOD2 and catalase in both aorta and liver tissues of atherosclerotic mice. CHWDT attenuated TNF-α and iNOS expression in RAW 264.7 cells, U937 cells and HUVECs, and restored eNOS expression in HUVECs. CHWDT decreased H2O2-induced cellular ROS generation in RAW 264.7 cells and U937 cells, and also decreased H2O2-induced mitochondrial ROS generation in RAW 264.7 cells. Furthermore, SOD1, SOD2 and catalase mRNA levels were increased by pre-treatment with CHWDT in H2O2 and LPS-stimulated RAW 264.7 cells, as well as in LPS-treated U937 and HUVECs. CHWDT not only decreased LPS-induced NF-κB p65 phosphorylation but also inhibited the translocation of p65 from the cytosol to the nucleus in RAW 264.7 macrophages. These results suggest that CHWDT exerts inhibitory effects on atherosclerosis-induced oxidative stress and inflammation via the NF-κB pathway.
Assuntos
Aterosclerose/prevenção & controle , Medicamentos de Ervas Chinesas/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , NF-kappa B/metabolismo , Tecido Adiposo/efeitos dos fármacos , Animais , Peso Corporal/efeitos dos fármacos , Citocinas/genética , Citocinas/metabolismo , Relação Dose-Resposta a Droga , Medicamentos de Ervas Chinesas/administração & dosagem , Humanos , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Masculino , Camundongos , Camundongos Knockout para ApoE , Estresse Oxidativo/efeitos dos fármacos , Células RAW 264.7 , Espécies Reativas de Oxigênio , Células U937RESUMO
The inflammatory cytokine tumor necrosis factor α (TNFα), upregulated in the obese condition, promotes protein degradation and is implicated in obesity-related skeletal muscle atrophy and age-related sarcopenia. Quercetin, a flavonoid, elicits antioxidative and anti-inflammatory activities. In this study, we investigated the effect of quercetin on TNFα-induced skeletal muscle atrophy as well as its potential mechanism of action. In this study, we observed that quercetin suppressed expression of TNFα-induced atrophic factors such as MAFbx/atrogin-1 and MuRF1 in myotubes, and it enhanced heme oxygenase-1 (HO-1) protein level accompanied by increased nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) in myotubes. The HO-1 inhibitor ZnPP suppressed the inhibitory actions of quercetin on TNFα-induced atrophic responses and degradation of IκB-α in myotubes. Moreover, quercetin supplementation to high-fat diet-fed obese mice inhibited obesity-induced atrophic responses in skeletal muscle, accompanied by upregulation of HO-1 and inactivation of nuclear factor-kappa B (NF-κB), and the quercetin actions were attenuated in Nrf2-deficient mice. These findings suggest that quercetin protects against TNFα-induced muscle atrophy under obese conditions through Nrf2-mediated HO-1 induction accompanied by inactivation of NF-κB. Quercetin may be used as a dietary supplement to protect against obesity-induced skeletal muscle atrophy.
Assuntos
Heme Oxigenase-1/genética , Atrofia Muscular/tratamento farmacológico , Atrofia Muscular/genética , Obesidade/complicações , Quercetina/administração & dosagem , Fator de Necrose Tumoral alfa/imunologia , Fator de Necrose Tumoral alfa/metabolismo , Animais , Heme Oxigenase-1/metabolismo , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fibras Musculares Esqueléticas/efeitos dos fármacos , Fibras Musculares Esqueléticas/metabolismo , Atrofia Muscular/etiologia , Atrofia Muscular/imunologia , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , Obesidade/genética , Obesidade/metabolismo , Fator de Necrose Tumoral alfa/genética , Regulação para CimaRESUMO
Obesity-induced hypothalamic inflammation is characterized by activation of microglia, which are resident macrophages of the central nervous system, and is implicated in the derangement of energy homeostasis, metabolic complications, and neurodegenerative diseases. Quercetin, a naturally occurring flavonoid, is known to protect against oxidative stress and inflammation-related metabolic complications. Here, we demonstrate that quercetin reduces obesity-induced hypothalamic inflammation by inhibiting microglia-mediated inflammatory responses, and the beneficial action of quercetin is associated with heme oxygenase (HO-1) induction. Quercetin markedly reduced the production of inflammatory mediators (monocyte chemoattractant protein (MCP)-1, interleukin (IL-6), IL-1ß, nitric oxide) by microglia stimulated with saturated fatty acid palmitate and/or lipid-laden microglia-conditioned medium. Quercetin also upregulated the expression of HO-1 in palmitate-treated lipid-laden microglia, and the actions of quercetin against microglia activation accompanied by IκBα degradation were abolished by a HO-1 inhibitor. Moreover, quercetin supplementation reduced the levels of inflammatory cytokines and microglia activation markers in the hypothalamus of high fat diet (HFD)-fed obese mice, which was accompanied by upregulation of HO-1. These findings indicate that quercetin suppresses microglia-mediated inflammatory responses via the induction of HO-1, and hence protects against obesity-induced hypothalamic inflammation.
Assuntos
Heme Oxigenase-1/metabolismo , Hipotálamo/patologia , Inflamação/induzido quimicamente , Proteínas de Membrana/metabolismo , Microglia/efeitos dos fármacos , Obesidade/complicações , Quercetina/farmacologia , Animais , Técnicas de Cultura de Células , Linhagem Celular , Meios de Cultivo Condicionados , Dieta Hiperlipídica/efeitos adversos , Gorduras na Dieta/administração & dosagem , Gorduras na Dieta/efeitos adversos , Regulação da Expressão Gênica/efeitos dos fármacos , Heme Oxigenase-1/genética , Inflamação/tratamento farmacológico , Masculino , Proteínas de Membrana/genética , Camundongos , Obesidade/induzido quimicamente , Distribuição AleatóriaRESUMO
High mobility group box 1 (HMGB1) is now recognized as a late mediator of sepsis. We tested hypothesis that ascorbic acid (AscA) induces heme oxygenase (HO)-1 which inhibits HMGB1 release in lipopolysaccharide (LPS)-stimulated cells and increases survival of septic mice. AscA increased HO-1 protein expression in a concentration- and time-dependent manner via Nrf2 activation in RAW 264.7 cells. HO-1 induction by AscA was significantly reduced by Nrf2 siRNA-transfected cells. Mutation of cysteine to serine of keap-1 proteins (C151S, C273S, and C288S) lost the ability of HO-1 induction by AscA, due to failure of translocation of Nrf-2 to nucleus. The PI3 kinase inhibitor, LY294002, inhibited HO-1 induction by AscA. Oxyhemoglobin (HbO2), LY294002, and ZnPPIX (HO-1 enzyme inhibitor) reversed effect of AscA on HMGB1 release. Most importantly, administration of AscA (200mg/kg, i.p.) significantly increased survival in LPS-induced endotoxemic mice. In cecal ligation and puncture (CLP)-induced septic mice, AscA reduced hepatic injury and serum HMGB1 and plasminogen activator inhibitor (PAI)-1 in a ZnPPIX-sensitive manner. In addition, AscA failed to increase survival in Nrf2 knockout mice by LPS. Thus, we concluded that high dose of AscA may be useful in the treatment of sepsis, at least, by activation of Nrf2/HO-1 signals.
Assuntos
Ácido Ascórbico/farmacologia , Proteína HMGB1/metabolismo , Heme Oxigenase-1/metabolismo , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Proteínas de Membrana/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Sepse/tratamento farmacológico , Transporte Ativo do Núcleo Celular , Animais , Ácido Ascórbico/uso terapêutico , Monóxido de Carbono/metabolismo , Linhagem Celular , Núcleo Celular/metabolismo , Chlorocebus aethiops , Ativação Enzimática , Fígado/efeitos dos fármacos , Fígado/patologia , Ativação de Macrófagos , Macrófagos/metabolismo , Masculino , Camundongos Endogâmicos ICR , NF-kappa B/metabolismo , Compostos Organometálicos/metabolismo , Compostos Organometálicos/farmacologia , Fosfatidilinositol 3-Quinases/metabolismo , Sepse/mortalidade , Sepse/fisiopatologiaRESUMO
Metabolic diseases, such as insulin resistance, type II diabetes, and obesity, are associated with a low-grade chronic inflammation (inflammatory stress), oxidative stress, and endoplasmic reticulum (ER) stress. Because the integration of these stresses is critical to the pathogenesis of metabolic diseases, agents and cellular molecules that can modulate these stress responses are emerging as potential targets for intervention and treatment of metabolic diseases. It has been recognized that heme oxygenase-1 (HO-1) plays an important role in cellular protection. Because HO-1 can reduce inflammatory stress, oxidative stress, and ER stress, in part by exerting antioxidant, anti-inflammatory, and antiapoptotic effects, HO-1 has been suggested to play important roles in pathogenesis of metabolic diseases. In the present review, we will explore our current understanding of the protective mechanisms of HO-1 in metabolic diseases and present some emerging therapeutic options for HO-1 expression in treating metabolic diseases, together with the therapeutic potential of curcumin and resveratrol analogues that have their ability to induce HO-1 expression.
Assuntos
Curcumina/análogos & derivados , Heme Oxigenase-1/biossíntese , Doenças Metabólicas/enzimologia , Estilbenos/metabolismo , Animais , Curcumina/farmacologia , Curcumina/uso terapêutico , Indução Enzimática/efeitos dos fármacos , Humanos , Doenças Metabólicas/tratamento farmacológico , Resveratrol , Estilbenos/farmacologia , Estilbenos/uso terapêuticoRESUMO
The Chung Hun Wha Dam Tang (CHWDT) herbal combination was reported to cease dizziness and phlegm. However, the effect of CHWDT in obesity has not yet been known mechanically. Therefore, we investigated whether this CHWDT could protect the cells from lipogenesis, gluconeogenesis, and inflammation in both in vivo and in vitro. CHWDT significantly decreased body weight, epididymal and perirenal fat content without affecting feed intake in high-fat diet-induced obese mice model. Additionally, CHWDT inhibited obesity-induced SREBP1, FAS, PGC1α, G6Pase, PEPCK and increased CPT1, ACO, and LCAD genes expression in vivo and in vitro. Proinflammatory cytokines like TNF-α and iNOS expression were reduced by CHWDT in both Raw264.7 macrophages and HepG2 cells. In addition, NO production was also significantly decreased by CHWDT in LPS-stimulated macrophages. Furthermore, AMPKα activation by CHWDT was involved in inhibition of obesity by reducing triglycerides production and increasing CPT1 expression. Based on all of the results, we suggest that CHWDT has inhibitory effects on obesity-induced lipogenesis, gluconeogenesis, and inflammation via AMPKα activation.
RESUMO
Salvia miltiorrhiza (Danshen), a traditional Chinese herbal medicine, is commonly used for the prevention and treatment of cardiovascular disorders including atherosclerosis. However, the mechanisms responsible for the vasoprotective effects of Danshen remain largely unknown. Salvianolic acid B (Sal B) represents one of the most bioactive compounds that can be extracted from the water-soluble fraction of Danshen. We investigated the effects of Danshen and Sal B on the inflammatory response in murine macrophages. Danshen and Sal B both induced the expression of heme oxygenase-1 (HO-1) and inhibited nitric oxide (NO) production and inducible NO synthase (iNOS) expression in lipopolysaccharide (LPS)-activated RAW 264.7 cells. Inhibition of HO activity using Sn-protoporphyrin-IX (SnPP) abolished the inhibitory effect of Sal B on NO production and iNOS expression. Sal B increased macrophage arginase activity in a dose-dependent manner and diminished LPS-inducible tumor necrosis factor-α production. These effects were also reversed by SnPP. These data suggest that HO-1 expression plays an intermediary role in the anti-inflammatory effects of Sal B. In contrast to the observations in macrophages, Sal B dose-dependently inhibited arginase activity in murine liver, kidney, and vascular tissue. Furthermore, Sal B increased NO production in isolated mouse aortas through the inhibition of arginase activity and reduction of reactive oxygen species production. We conclude that Sal B improves vascular function by inhibiting inflammatory responses and promoting endothelium-dependent vasodilation. Taken together, we suggest that Sal B may represent a potent candidate therapeutic for the treatment of cardiovascular diseases associated with endothelial dysfunction.
Assuntos
Benzofuranos/farmacologia , Medicamentos de Ervas Chinesas/farmacologia , Heme Oxigenase-1/metabolismo , Macrófagos/efeitos dos fármacos , Óxido Nítrico Sintase Tipo II/antagonistas & inibidores , Óxido Nítrico/metabolismo , Animais , Arginase/metabolismo , Western Blotting , Ensaio de Desvio de Mobilidade Eletroforética , Fibrinolíticos/farmacologia , Humanos , Lipopolissacarídeos/farmacologia , Macrófagos/enzimologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fenantrolinas/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Salvia miltiorrhiza/química , Fator de Necrose Tumoral alfa/metabolismo , Doenças Vasculares/prevenção & controleRESUMO
Excess production of nitric oxide by activated macrophages via inducible nitric oxide synthase leads to the development of various inflammatory diseases. Heme oxygenase-1 expression via activation of nuclear factor-erythroid 2-related factor 2 inhibits nitric oxide production and inducible nitric oxide synthase expression in activated macrophages. Okanin is one of the most abundant chalcones found in the genus Bidens (Asteraceae) that is used as various folk medications in Korea and China for treating inflammation. Here, we found that okanin (possessing the α-ß unsaturated carbonyl group) induced heme oxygenase-1 expression via nuclear factor-erythroid 2-related factor 2 activation in RAW264.7 macrophages. 3-Penten-2-one, of which structure, as in okanin, possesses the α-ß unsaturated carbonyl group, also induced nuclear factor-erythroid 2-related factor 2-dependent heme oxygenase-1 expression, while both 2-pentanone (lacking a double bond) and 2-pentene (lacking a carbonyl group) were virtually inactive. In lipopolysaccharide-activated RAW264.7 macrophages, both okanin and 3-penten-2-one inhibited nitric oxide production and inducible nitric oxide synthase expression via heme oxygenase-1 expression. Collectively, our findings suggest that by virtue of its α-ß unsaturated carbonyl functional group, okanin can inhibit nitric oxide production and inducible nitric oxide synthase expression via nuclear factor-erythroid 2-related factor 2-dependent heme oxygenase-1 expression in lipopolysaccharide-activated macrophages.
RESUMO
Cudratricusxantone A (CTXA), isolated from the roots of Cudrania tricuspidata Bureau (Moraceae), has potent hepatoprotective, antiproliferative, and monoamine oxidase inhibitory effects. In this study, we examined whether CTXA could protect HT22-immortalized hippocampal cells against glutamate-induced oxidative stress through the induction of heme oxygenase (HO)-1 expression. CTXA induced the expression of HO-1 and increased HO activity dose- and time-dependently. CTXA also suppressed glutamate-induced ROS generation in HT22 cells. Interestingly, treatment of neuronal cells with CTXA enhanced cellular resistance to glutamate oxidative stress. The protective effect of CTXA was abrogated by tin protoporphyrin IX, an HO inhibitor. In addition, treatment with the HO-1 inducer, cobalt protoporphyrin IX, and bilirubin, one of the enzymatic products of HO-1, produced comparable protection. These results demonstrate that CTXA protects neuronal cells from glutamate-induced oxidative stress via the induction of HO-1.
Assuntos
Heme Oxigenase-1/metabolismo , Hipocampo/efeitos dos fármacos , Moraceae/química , Síndromes Neurotóxicas/prevenção & controle , Xantonas/uso terapêutico , Animais , Linhagem Celular , Ácido Glutâmico/toxicidade , Hipocampo/enzimologia , Camundongos , Casca de Planta/química , Raízes de Plantas/química , Xantonas/isolamento & purificação , Xantonas/farmacologiaRESUMO
Doxorubicin (Dox) is a highly effective anticancer drug but exhibits cumulative dose-dependent cardiomyopathy. In this study, we investigated effects of Magnolia seed extract (MagS) on the Dox-induced cardiotoxicity. The results showed that MagS significantly reduces doxorubicin (Dox)-induced increase in intracellular Ca2+ concentration ([Ca2+]i), generation of reactive oxygen species (ROS), and apoptosis in rat cardiomyocytes. Analyses of the bioactive compounds in MagS by thin layer chromatography and gas chromatography/mass spectroscopy revealed that bioactive compounds in MagS are linoleic acid, oleic acid, and palmitic acid. All three fatty acids were able to inhibit the Dox-induced increase in [Ca2+]i, ROS generation, and apoptosis with a similar potency. Efficacy of MagS was examined in in vivo using a murine Dox-induced cardiomyopathy model. Dox (12 mg/kg, intravenously) was administered to mice and treated with the MagS (2 mg/kg/d, intraperitoneally) or saline for three weeks. Dox-treated mice showed structural disarray in heart tissue, including lymphocyte infiltration and loss of body weight. In contrast, treatment of the MagS substantially attenuated the Dox-induced cardiac damages including the loss of body weight. These results indicate that fatty acids in MagS and other seeds may ameliorate cardiotoxicity of the anticancer drug.
Assuntos
Antibióticos Antineoplásicos/antagonistas & inibidores , Antibióticos Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Cálcio/metabolismo , Doxorrubicina/antagonistas & inibidores , Doxorrubicina/farmacologia , Ácidos Graxos/farmacologia , Miócitos Cardíacos/efeitos dos fármacos , Animais , Peso Corporal/efeitos dos fármacos , Caspase 3/metabolismo , Cromatografia em Camada Fina , Ativação Enzimática/efeitos dos fármacos , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Magnolia/química , Camundongos , Camundongos Endogâmicos BALB C , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Sementes/química , Espectrofotometria UltravioletaRESUMO
Inducible heme oxygenase (HO)-1 is known to play a major role in the pathogenesis of several diseases, and it protects cells against oxidant-mediated injury. The bioassay-guided fractionation of the EtOH extract of the flowered fruit-spike of Prunella vulgaris L. (Labiatae) yielded two ursane-type triterpenes, 3beta,23-dihydroxyurs-12-en-28-oic acid (23-hydroxyursolic acid) (1) and 3beta-hydroxyurs-12-en-28-oic acid (ursolic acid) (2). Western blotting demonstrated that treatment with compound 1 increased the expression of HO-1 in a dose-dependent manner in human liver-derived HepG2 cells. Investigation of structure-related HO-1 inducing activity suggested that the hydroxyl group at the C-23 position in the ursane skeleton is important for this activity.
Assuntos
Heme Oxigenase-1/biossíntese , Extratos Vegetais/farmacologia , Prunella/química , Western Blotting , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Indução Enzimática , Flores/química , Frutas/química , Hepatócitos/efeitos dos fármacos , Hepatócitos/enzimologia , Humanos , Estrutura Molecular , Extratos Vegetais/efeitos adversos , Extratos Vegetais/isolamento & purificação , Pós , Relação Estrutura-AtividadeRESUMO
Piperine is a major component of black pepper, Piper nigrum Linn, used widely in traditional medicine. In this study, we examined whether piperine could protect House Ear Institute-Organ of Corti 1 (HEI-OC1) cells against cisplatin-induced apoptosis through the induction of heme oxygenase (HO)-1 expression. Piperine (10-100 microM) induced the expression of HO-1 in dose- and time-dependent manners. Piperine also induced antioxidant response element-luciferase and translocated nuclear factor-E2-related factor-2 (Nrf2) to nucleus. Piperine activated the c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase and p38 mitogen-activated protein kinase (MAPK) pathways, and the JNK pathway played an important role in piperine-induced HO-1 expression. Piperine protected the cells against cisplatin-induced apoptosis. The protective effect of piperine was abrogated by zinc protoporphyrin IX, an HO inhibitor, and antisense oligodeoxynucleotides against HO-1 gene. These results demonstrate that the expression of HO-1 by piperine is mediated by both JNK pathway and Nrf2, and the expression inhibits cisplatin-induced apoptosis in HEI-OC1 cells.
Assuntos
Alcaloides/farmacologia , Antineoplásicos/toxicidade , Apoptose/efeitos dos fármacos , Benzodioxóis/farmacologia , Cisplatino/toxicidade , Heme Oxigenase-1/metabolismo , Órgão Espiral/efeitos dos fármacos , Piperidinas/farmacologia , Alcamidas Poli-Insaturadas/farmacologia , Animais , Núcleo Celular/metabolismo , Células Cultivadas , Relação Dose-Resposta a Droga , Células Ciliadas Auditivas Externas/metabolismo , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Camundongos , Fator 2 Relacionado a NF-E2/metabolismo , Órgão Espiral/enzimologia , Transdução de Sinais , TransfecçãoRESUMO
Inducible heme oxygenase (HO)-1 acts against oxidants that are thought to play a major role in the pathogenesis of several diseases. The alpha-methylene-gamma-butyrolactone (CH2-BL) structural unit, which characterizes a group of naturally occurring sesquiterpene lactones, is known to possess numerous biological activities. In the present study, we evaluated dehydrocostus lactone possessing CH2-BL moiety, one of the bioactive constituents of the medicinal plant Saussurea lappa, as an inducer of cytoprotective HO-1. In HepG2 cells, treatment with dehydrocostus lactone induced HO-1 expression and increased HO activity in a concentration-dependent manner. Similar results were also observed when the cells were incubated with CH2-BL, a parent structure of dehydrocostus lactone. In contrast, mokko lactone, a reduced product of dehydrocostus lactone, and alpha-methyl-gamma-butyrolactone (CH3-BL), a parent structure of mokko lactone, did not induce HO-1 expression. Pretreatment with either dehydrocostus lactone or CH2-BL for 6 h protected the cells from hydrogen peroxide-mediated toxicity, whereas mokko lactone or CH3-BL failed to exert a cytoprotective action. Inhibition of HO-1 expression by HO-1 small interfering RNA (siRNA) abrogated cellular protection afforded by dehydrocostus lactone or CH2-BL. In addition, dehydrocostus lactone caused the nuclear accumulation of the nuclear factor E2-related factor 2 (Nrf2) and increased the promoter activity of antioxidant response element (ARE). Using Nrf2 siRNA, Nrf2 activation was confirmed to contribute to cytoprotective HO-1 expression by dehydrocostus lactone or CH2-BL. Collectively, our findings suggest that CH2-BL moiety in dehydrocostus lactone increases cellular resistance to oxidant injury in HepG2 cells, presumably through Nrf2/ARE-dependent HO-1 expression.
Assuntos
4-Butirolactona/análogos & derivados , Antioxidantes/farmacologia , Citoproteção , Heme Oxigenase-1/fisiologia , Lactonas/farmacologia , Fator 2 Relacionado a NF-E2/fisiologia , Sesquiterpenos/farmacologia , 4-Butirolactona/farmacologia , Transporte Ativo do Núcleo Celular , Linhagem Celular , Heme Oxigenase-1/genética , Humanos , Lactonas/química , Elementos de Resposta/fisiologia , Sesquiterpenos/químicaRESUMO
BACKGROUND: Mite antigen, extract from Dermatophagoides farinae in house dust, is a well-known causative agent of atopy or allergic diseases, which involves many inflammatory cytokines/chemokines expression. Heme oxygenase 1 (HO1) has recently emerged as an important cytoprotective enzyme against oxidative stress and inflammatory responses in many cell types. OBJECTIVE: The aim of this study was to investigate the possible mechanism by which wogonin, a natural product isolated from Scutellaria baicalensis, inhibited the mite antigen-induced chemokine expression in human keratinocytes, HaCaT cells. METHODS: The level of chemokine expression was measured by reverse transcription-polymerase chain reaction (RT-PCR) and signaling study was performed by Western blot analysis. RESULTS: The mite antigen-induced thymus- and activation-regulated chemokine (TARC/CCL17) expression in a dose-dependent manner via extracellular signal-regulated kinase (ERK) activation. However, it did not affect the expression of other chemokines including macrophage-derived chemokine (MDC/CCL22), RANTES, and IL-8. Interestingly, wogonin significantly suppressed the mite antigen-induced TARC expression via the induction of HO1. This suppression was completely restored by HO1 siRNA, suggesting a direct role of HO1 for the suppressive effect. Furthermore, exogenous CO, but not other end products of HO1 activity, also suppressed the mite antigen-induced TARC expression. CONCLUSION: Wogonin induces HO1 expression, which in turn HO1 and/or CO suppresses TARC expression induced by mite antigen in human HaCaT cells.
Assuntos
Quimiocinas CC/genética , Dermatite Atópica/tratamento farmacológico , Flavanonas/farmacologia , Heme Oxigenase-1/genética , Queratinócitos/efeitos dos fármacos , Queratinócitos/imunologia , Animais , Antígenos de Dermatophagoides/imunologia , Antígenos de Dermatophagoides/farmacologia , Linhagem Celular Transformada , Quimiocina CCL17 , Dermatite Atópica/imunologia , Dermatophagoides farinae , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/farmacologia , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Flavanonas/química , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/imunologia , Heme Oxigenase-1/metabolismo , Humanos , Técnicas In Vitro , Queratinócitos/citologia , Fosforilação/efeitos dos fármacos , Scutellaria baicalensisRESUMO
The mushroom Phellinus linteus (PL) has been shown to have antitumor and immunostimulatory effects. We hypothesized that the hot water extract of PL (WEPL) exerts its significant immunostimulatory effect by inducing production of the Th1-derived cytokine interferon-gamma (IFN-gamma) by T lymphocytes. T lymphocytes were isolated from the mice fed with 200 mg/kg of WEPL once a day for 4 weeks and then stimulated with the mitogen concanavaline A (Con A). IFN-gamma gene and intracellular protein expressions were analyzed by RT-PCR and flow cytometry, respectively. The production of IFN-gamma was measured by enzyme-linked immunosorbent assay. WEPL significantly enhanced the transcription of IFN-gamma mRNA. The effect of WEPL on IFN-gamma expression was further supported by a concomitant increase in the number of cells with intracellular IFN-gamma protein as well as the secretion of IFN-gamma. However, WEPL did not modulate either gene expression or protein secretion of interleukin-4, a Th2-associated cytokine, by Con A-stimulated T lymphocytes. Our results demonstrate that one of the potentially beneficial antitumor and immunostimulatory effects of WEPL may be mediated through the enhancement of IFN-gamma secretion by T lymphocytes.
Assuntos
Adjuvantes Imunológicos/administração & dosagem , Basidiomycota , Misturas Complexas/administração & dosagem , Citocinas/biossíntese , Regulação da Expressão Gênica/efeitos dos fármacos , Células Th1/metabolismo , Células Th2/metabolismo , Adjuvantes Imunológicos/química , Administração Oral , Animais , Basidiomycota/química , Células Cultivadas , Misturas Complexas/química , Citocinas/imunologia , Avaliação Pré-Clínica de Medicamentos , Feminino , Regulação da Expressão Gênica/imunologia , Técnicas In Vitro , Masculino , Camundongos , Células Th1/citologia , Células Th1/imunologia , Células Th2/citologia , Células Th2/imunologiaRESUMO
Scoparone is a major component of the shoot of Artemisia capillaris (Compositae), which has been used for the treatment of hepatitis and biliary tract infection in oriental countries. In the present study we observed that, scorparone exhibited no cytotoxic effect in unstimulated macrophages, but reduced the release of nitric oxide (NO) and prostaglandin E2 (PGE2) upon stimulation by IFN-gamma/LPS or LPS. The inhibitory effects were found to be in conjuction with the suppression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in IFN-gamma/LPS stimulated RAW 264.7 cells. Moreover, scoparone also attenuated the production of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta and IL-6 in LPS-stimulated RAW264.7 cells. These results suggest that scoparone decreases the production of the inflammatory mediators such as NO and PGE2 in macrophages by inhibiting iNOS and COX-2 expression.
Assuntos
Anti-Inflamatórios/farmacologia , Artemisia/química , Cumarínicos/farmacologia , Mediadores da Inflamação/metabolismo , Interferon gama/farmacologia , Lipopolissacarídeos/farmacologia , Macrófagos/metabolismo , Animais , Anti-Inflamatórios/isolamento & purificação , Western Blotting , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Cumarínicos/isolamento & purificação , Ciclo-Oxigenase 2 , Citocinas/biossíntese , Dinoprostona/biossíntese , Macrófagos/efeitos dos fármacos , Camundongos , Óxido Nítrico/biossíntese , Óxido Nítrico Sintase/biossíntese , Óxido Nítrico Sintase Tipo II , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Prostaglandina-Endoperóxido Sintases/metabolismo , Estimulação Química , Sais de Tetrazólio , TiazóisRESUMO
We studied the effect of 4-acetyl-12,13-epoxyl-9-trichothecene-3, 15-diol (AETD) isolated from Isaria japonica, one of the most popular Chinese fungal medicines, on the induction of apoptosis in rat bladder carcinoma NBT-II cells. AETD was cytotoxic to NBT-II cells, and this cytotoxic effect appears to be attributed to its induction of apoptotic cell death, as AETD induced nuclear morphological changes and internucleosomal DNA fragmentation, and increased the proportion of hypodiploid cells and activity of caspase-3. AETD treatment also decreased the expression of the anti-apoptotic protein Bcl-2 and increased the expression of the pro-apoptotic protein Bax. These results provide important information in understanding the mechanism(s) of AETD-induced apoptosis.
Assuntos
Apoptose/efeitos dos fármacos , Carcinoma/patologia , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Tricotecenos/farmacologia , Neoplasias da Bexiga Urinária/patologia , Animais , Carcinoma/metabolismo , Caspase 3 , Caspases/metabolismo , Linhagem Celular Tumoral/efeitos dos fármacos , Fragmentação do DNA , DNA de Neoplasias/análise , Microscopia de Fluorescência , Ratos , Neoplasias da Bexiga Urinária/metabolismo , Proteína X Associada a bcl-2RESUMO
Farnesylation of p21(ras) is an important step in the intracellular signaling pathway of growth factors, hormones, and immune stimulants. We synthesized a potent and selective farnesyltransferase inhibitor (LB42708) with IC(50) values of 0.8 nM in vitro and 8 nM in cultured cells against p21(ras) farnesylation and examined the effects of this inhibitor in the settings of inflammation and arthritis. LB42708 suppressed NF-kappaB activation and iNOS promoter activity by suppressing the I-kappaB kinase activity and I-kappaBalpha degradation. The inhibitor suppressed the expression of inducible NO synthase, cyclooxygenase-2, TNF-alpha, and IL-1beta and the production of NO and PGE(2) in immune-activated macrophages and osteoblasts as well as LPS-administrated mice. Furthermore, in vivo administration of LB42708 significantly decreased the incidence and severity of arthritis as well as mRNA expression of inducible NO synthase, cyclooxygenase-2, TNF-alpha, and IL-1beta in the paws of collagen-induced arthritic mice compared with controls. These observations indicate that the anti-inflammatory and antiarthritic effects of the farnesyltransferase inhibitor may be ascribed to the inhibition of I-kappaB kinase activity and subsequent suppression of NF-kappaB-dependent inflammatory gene expression through the suppression of p21(ras) farnesylation. Together, these findings reveal that the inhibitory effect of LB42708 on p21(ras)-dependent NF-kappaB activation may have potential therapeutic value for arthritis and other inflammatory diseases.