RESUMEN
Diesel exhaust particles (DEPs) are a major cause of cancer progression as well as a variety of acute and chronic diseases. It is well-known that programmed death-ligand 1 (PD-L1) is an immune checkpoint molecule that can induce immune escape in tumor cells. However, the function of PD-L1 in bronchial epithelial cells or how PD-L1 relates to cellular oxidation under DEPs-mediated oxidative stress is not well known. In this study, we investigated how PD-L1 affected DEPs-induced oxidative stress and cytotoxicity in human bronchial epithelial (HBE) cells, Beas-2B. DEPs not only induced intracellular reactive oxygen species (ROS) production, but also increased PD-L1 expression in HBE cells. Beas-2B cells overexpressing PD-L1 showed higher levels of ROS production, DNA damage, and apoptosis after DEPs treatment compared to control cells. In particular, the expression of an antioxidant enzyme heme-oxygenase-1 (HO-1) and nuclear translocation and transcriptional activity of Nrf2, a major regulator of HO-1, were lower in Beas-2B overexpressing PD-L1 cells than in control cells. DEPs-induced ROS generation, DNA damage and apoptosis in Beas-2B cells overexpressing PD-L1 were significantly restored by overexpressing HO-1. Collectively, our results suggest that DEPs can increase the expression of PD-L1 in HBE cells and that overexpressing PD-L1 might eventually promote DEPs-induced oxidative DNA damage and apoptosis.
Asunto(s)
Antígeno B7-H1 , Emisiones de Vehículos , Humanos , Emisiones de Vehículos/toxicidad , Antígeno B7-H1/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Estrés Oxidativo , Células Epiteliales/patologíaRESUMEN
Toll-like receptors (TLRs) are integral membrane-bound receptors that are central to innate and adaptive immune responses. They are known to activate a cascade of downstream signals to induce the secretion of inflammatory cytokines, chemokines, and type I interferons. Dysregulated activation of TLR signaling pathways can induce the activation of various transcription factors, such as nuclear factor kappa B (NF-κB) and interferon regulatory factor 3 (IRF3). TLRs act via MyD88- and TRIF-mediated pathways to induce inflammatory responses. To evaluate the therapeutic potential of isobavachalcone (IBC), a natural chalcone component of Angelica keiskei, we examined its effects on signal transduction via TLR signaling pathways. IBC inhibited the activation of NF-κB and IRF3 induced by TLR agonists and their target genes. IBC also inhibited the activation of NF-κB and IRF3 induced by overexpression of downstream signaling components of TLR signaling pathways. These results suggest that IBC can regulate both MyD88- and TRIF-dependent signaling pathways of TLRs, resulting in a dramatic increase of new therapeutic options for various inflammatory diseases involving TLRs.
Asunto(s)
Chalconas , Proteínas Adaptadoras del Transporte Vesicular/metabolismo , Proteínas Adaptadoras del Transporte Vesicular/farmacología , Chalconas/farmacología , Factor 88 de Diferenciación Mieloide/metabolismo , Factor 88 de Diferenciación Mieloide/farmacología , FN-kappa B , Transducción de Señal , Relación Estructura-Actividad , Receptores Toll-Like/agonistas , Receptores Toll-Like/metabolismoRESUMEN
Sauchinone, a lignan isolated from Saururus chinenesis, is known to exhibit anti-inflammatory and anti-oxidant effects. Recently, sauchinone has been reported to inhibit the growth of various cancer cells, but its effects on breast cancer cells remain poorly understood. In the present study, we investigated the effects of sauchinone on the growth of breast cancer cells along with the underlying molecular mechanisms. Our results show that sauchinone treatment markedly inhibited the proliferation, migration, and invasion of breast cancer cells. Sauchinone reduced the phosphorylation of Akt, ERK, and CREB increased by transforming growth factor-ß (TGF-ß). In particular, sauchinone treatment suppressed the expression of matrix metalloproteinase (MMP)-13 (MMP13) by regulating the Akt-CREB signaling pathway. Sauchinone was less effective in inhibiting cell migration in Mmp13-knockdown cells than in control cells, suggesting that MMP13 may be a novel target for sauchinone. Our study suggests that sauchinone inhibits the growth of breast cancer cells by attenuating the Akt-CREB-MMP13 pathway. In addition, the targeted inhibition of MMP13 by sauchinone represents a promising approach for the treatment of breast cancer.
Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Benzopiranos/farmacología , Neoplasias de la Mama/tratamiento farmacológico , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Dioxoles/farmacología , Metaloproteinasa 13 de la Matriz/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Neoplasias de la Mama/enzimología , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Línea Celular Tumoral , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Metaloproteinasa 13 de la Matriz/genética , Invasividad Neoplásica , Fosforilación , Transducción de SeñalRESUMEN
Toll-like receptors (TLRs) can recognize specific signatures of invading microbial pathogens and activate a cascade of downstream signals to induce the secretion of inflammatory cytokines, chemokines, and type I interferons. The activation of TLRs triggers two downstream signaling pathways: the MyD88- and the TRIF-dependent pathways. To evaluate the therapeutic potential of epoxomicin, a member of the linear peptide α',ß'-epoxyketone first isolated from an actinomycetes strain, we examined its effects on signal transduction via TLR signaling pathways. Epoxomicin inhibited the activation of NF-kB and IRF3 induced by TLR agonists, decreased the expression of interferon-inducible protein-10, and inhibited the activation of NF-kB and IRF3 induced by overexpression of downstream signaling components of TLR signaling pathways. These results suggest that epoxomicin can regulate both the MyD88- and TRIF-dependent signaling pathways of TLRs. Thus, it might have potential as a new therapeutic drug for a variety of inflammatory diseases.
Asunto(s)
Transducción de Señal/efectos de los fármacos , Receptores Toll-Like/metabolismo , Animales , Células HEK293 , Humanos , Factor 3 Regulador del Interferón/metabolismo , Ratones , Factor 88 de Diferenciación Mieloide/metabolismo , FN-kappa B/metabolismo , Oligopéptidos/farmacología , Células RAW 264.7RESUMEN
Gremlin-1 (GREM1), one of the antagonists of bone morphogenetic proteins (BMPs), has recently been reported to be overexpressed in a variety of cancers including breast cancer. GREM1 is involved in tumor promotion, but little is known about its role in the glycolysis of cancer cells. In this study, we investigated the role of GREM1 in glycolysis of breast cancer cells and its underlying molecular mechanisms. We first observed that glucose uptake and lactate production were increased in GREM1-overexpressing breast cancer cells. GREM1 increased the expression of hexokinase-2 (HK2), which catalyzes the phosphorylation of glucose, the first step in glycolysis. In addition, GREM1 activated STAT3 transcription factor through the ROS-Akt signaling pathway. The ROS-Akt-STAT3 axis activated by GREM1 was involved in promoting glucose uptake by increasing the expression of HK2 in breast cancer cells. Therefore, our study suggested a new mechanism by which GREM1 is involved in breast cancer promotion by increasing glycolysis in breast cancer cells.
Asunto(s)
Neoplasias de la Mama/metabolismo , Glucólisis/fisiología , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Factor de Transcripción STAT3/metabolismo , Transducción de Señal , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Línea Celular Tumoral , Femenino , Regulación Neoplásica de la Expresión Génica , Glucosa/metabolismo , Hexoquinasa/genética , Humanos , Péptidos y Proteínas de Señalización Intercelular/genética , Ácido Láctico/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Toll-like receptors (TLRs) are a group of pattern-recognition receptors (PRRs) that are at the core of innate and adaptive immune responses. TLRs activation triggers the activation of two downstream signaling pathways, the myeloid differential factor 88 (MyD88)- and toll-interleukin-1 receptor domain-containing adapter inducing interferon-ß (TRIF)-dependent pathways. To evaluate the therapeutic potential of DHL, a natural sesquiterpene lactone derived from Inulahelenium L. and Saussurea lappa, we examined its effect on signal transduction via the TLR signaling pathways. DHL inhibited the activation of nuclear factor-κB (NF-κB) and interferon regulatory factor 3 (IRF3), the representative transcription factors involved in the inflammatory response, induced by TLR agonists, as well as the expression of cyclooxygenase-2 and interferon inducible protein-10. DHL also inhibited the activation of NF-κB and IRF3 induced by the overexpression of downstream signaling components of the TLRs signaling pathways. All results suggest that DHL might become a new therapeutic drug for a variety of inflammatory diseases.
Asunto(s)
Antiinflamatorios/farmacología , Inflamación/tratamiento farmacológico , Lactonas/farmacología , Sesquiterpenos/farmacología , Transducción de Señal/efectos de los fármacos , Receptores Toll-Like/metabolismo , Proteínas Adaptadoras del Transporte Vesicular/metabolismo , Animales , Antiinflamatorios/uso terapéutico , Células HEK293 , Humanos , Inflamación/inmunología , Factor 3 Regulador del Interferón/metabolismo , Inula/química , Lactonas/uso terapéutico , Ratones , Factor 88 de Diferenciación Mieloide/inmunología , Factor 88 de Diferenciación Mieloide/metabolismo , FN-kappa B/metabolismo , Células RAW 264.7 , Saussurea/química , Sesquiterpenos/uso terapéutico , Transducción de Señal/inmunologíaRESUMEN
Toll-like receptors (TLRs) play a crucial role in the induction of innate immune response against bacterial and viral infections. TLRs induce downstream signaling via MyD88- and TRIF-dependent pathways. Cardamonin is a naturally occurring chalcone from Alpinia species exhibiting anti-inflammatory effects. However, the principal molecular mechanisms remain unclear. The objective of this study was to investigate the role of cardamonin in TLR signaling pathways. Cardamonin inhibited NF-κB activation as well as COX-2 expression induced by TLR agonists. Cardamonin inhibited the activation of IRF3 and the expression of interferon-inducible protein-10 (IP-10) induced by TLR3 or TLR4 agonists. Cardamonin also inhibited ligand-independent NF-κB activation overexpressed by MyD88, IKKß, or p65 and IRF3 activation overexpressed by TRIF, TBK1, or IRF3. However, cardamonin had no effect on TBK1 kinase activity in vitro. These results suggest that cardamonin modulates both the MyD88- and TRIF-dependent pathways of TLRs and represents a potentially new anti-inflammatory candidate.
Asunto(s)
Proteínas Adaptadoras del Transporte Vesicular/fisiología , Chalconas/farmacología , Factor 88 de Diferenciación Mieloide/fisiología , Transducción de Señal/efectos de los fármacos , Receptores Toll-Like/fisiología , Proteínas Adaptadoras del Transporte Vesicular/antagonistas & inhibidores , Animales , Factor 3 Regulador del Interferón/fisiología , Ratones , FN-kappa B/antagonistas & inhibidores , Células RAW 264.7RESUMEN
Toll-like receptors (TLRs) play a crucial role in danger recognition and induction of innate immune response against bacterial and viral infections. The TLR adaptor molecule, toll-interleukin-1 receptor domain-containing adapter inducing interferon-ß (TRIF), facilitates TLR3 and TLR4 signaling, leading to the activation of the transcription factor, NF-κB and interferon regulatory factor 3 (IRF3). Andrographolide, the active component of Andrographis paniculata, exerts anti-inflammatory effects; however, the principal molecular mechanisms remain unclear. The objective of this study was to investigate the role of andrographolide in TLR signaling pathways. Andrographolide suppressed NF-κB activation as well as COX-2 expression induced by TLR3 or TLR4 agonists. Andrographolide also suppressed the activation of IRF3 and the expression of interferon inducible protein-10 (IP-10) induced by TLR3 or TLR4 agonists. Andrographolide attenuated ligand-independent activation of IRF3 following overexpression of TRIF, TBK1, or IRF3. Furthermore, andrographolide inhibited TBK1 kinase activity in vitro. These results indicate that andrographolide modulates the TRIF-dependent pathway of TLRs by targeting TBK1 and represents a potential new anti-inflammatory candidate.
Asunto(s)
Antiinflamatorios/uso terapéutico , Diterpenos/uso terapéutico , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Adaptadoras del Transporte Vesicular/metabolismo , Andrographis/inmunología , Animales , Quimiocina CXCL10/metabolismo , Factor 3 Regulador del Interferón/metabolismo , Ratones , FN-kappa B/metabolismo , Células RAW 264.7 , Transducción de Señal , Receptor Toll-Like 3/metabolismo , Receptor Toll-Like 4/metabolismo , Activación TranscripcionalRESUMEN
Toll-like receptor 4 (TLR4) recognizes lipopolysaccharide (LPS) and triggers the activation of myeloid differention factor 88 (MyD88) and the Toll/interleukin-1 receptor domain-containing adapter, inducing interferon-ß (TRIF)-dependent major downstream signaling pathways. To evaluate the therapeutic potential of 1-[5-methoxy-2-(2-nitrovinyl)phenyl]pyrrolidine (MNP), previously synthesized in our laboratory, its effect on signal transduction via the TLR signaling pathways was examined. Here, we investigated whether MNP modulates the TLR4 signaling pathways and which anti-inflammatory target in TLR4 signaling is regulated by MNP. MNP inhibited the activation of nuclear factor-κB (NF-κB) induced by LPS (TLR4 agonist), and it also inhibited the expression of cyclooxygenase-2 and inducible nitric oxide synthase. MNP inhibited LPS-induced NF-κB activation by targeting TLR4 dimerization in addition to IKKß. These results suggest that MNP can modulate the TLR4 signaling pathway at the receptor level to decrease inflammatory gene expression.
Asunto(s)
Nitrocompuestos/farmacología , Multimerización de Proteína/efectos de los fármacos , Pirrolidinas/farmacología , Receptor Toll-Like 4/metabolismo , Animales , Antiinflamatorios/farmacología , Células Cultivadas , Ciclooxigenasa 2/biosíntesis , Relación Dosis-Respuesta a Droga , Quinasa I-kappa B/antagonistas & inhibidores , Lipopolisacáridos , Ratones , FN-kappa B/biosíntesis , Óxido Nítrico Sintasa de Tipo II/biosíntesis , Transducción de Señal/efectos de los fármacosRESUMEN
Toll-like receptors (TLRs) play significant roles in recognizing the pathogen-associated molecular patterns that induce innate immunity, and subsequently, acquired immunity. In general, TLRs have two downstream signaling pathways, the myeloid differential factor 88 (MyD88)-dependent and toll-interleukin-1 receptor domain-containing adapter-inducing interferon-ß (TRIF)-dependent pathways, which lead to the activation of nuclear factor-kappa B (NF-κB) and interferon regulatory factor 3 (IRF3). 1-[5-methoxy-2-(2-nitrovinyl)phenyl]pyrrolidine (MNP) has been previously synthesized in our laboratory. To evaluate the therapeutic potential of MNP, its effect on signal transduction via the TLR signaling pathways was examined. MNP was shown to inhibit the activation of NF-κB and IRF3 induced by TLR agonists, as well as to inhibit the expression of cyclooxygenase-2, inducible nitric oxide synthase, and interferon inducible protein-10. MNP also inhibited the activation of NF-κB and IRF3 induced by the overexpression of downstream signaling components of the MyD88- or TRIF-dependent signaling pathways. These results suggest that MNP can modulate MyD88- and TRIF-dependent signaling pathways of TLRs, leading to decreased inflammatory gene expression.
Asunto(s)
Nitrocompuestos/farmacología , Pirrolidinas/farmacología , Receptores Toll-Like/agonistas , Proteínas Adaptadoras del Transporte Vesicular/metabolismo , Animales , Quimiocina CXCL10/metabolismo , Ciclooxigenasa 2/metabolismo , Células HEK293 , Humanos , Inmunidad Innata , Factor 3 Regulador del Interferón/genética , Factor 3 Regulador del Interferón/metabolismo , Ratones , Factor 88 de Diferenciación Mieloide/metabolismo , FN-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Nitrocompuestos/química , Pirrolidinas/química , Células RAW 264.7 , Transducción de Señal/efectos de los fármacosRESUMEN
Toll-like receptor 4 (TLR4) recognizes LPS and triggers the activation of the myeloid differential factor 88 (MyD88)- and toll-interleukin-1 receptor domain-containing adapter, inducing interferon-ß (TRIF)-dependent major downstream signaling pathways. Previously, we presented biochemical evidence that 1-[4-Fluoro-2-(2-nitrovinyl)phenyl]pyrrolidine (FPP), which was synthesized in our laboratory, inhibits NF-κB activation induced by LPS. Here, we investigated whether FPP modulates the TLR4 downstream signaling pathways and what anti-inflammatory target in TLR4 signaling is regulated by FPP. FPP inhibited LPS-induced NF-κB activation by targeting TLR4 dimerization. These results suggest that FPP can modulate the TLR4 signaling pathway at the receptor level to decrease inflammatory gene expression.
Asunto(s)
Lipopolisacáridos/farmacología , Multimerización de Proteína/efectos de los fármacos , Pirrolidinas/farmacología , Receptor Toll-Like 4/metabolismo , Compuestos de Vinilo/farmacología , Animales , Células Cultivadas , Células HEK293 , Humanos , Ratones , Estructura Molecular , FN-kappa B/metabolismo , Unión Proteica/efectos de los fármacos , Receptor Toll-Like 4/químicaRESUMEN
Toll-like receptors (TLRs) recognize distinct pathogen-associated molecular patterns and play a critical role in innate immune responses. TLR signaling pathways can be largely classified as either myeloid differential factor 88 (MyD88)- or toll-interleukin-1 receptor domain-containing adapter inducing interferon-ß (TRIF)-dependent pathways. Compound of Designation red 10 binding (CDr10b) was synthesized to investigate its role in neuroinflammatory diseases. This study was conducted to determine whether CDr10b can affect TLR signaling pathways. CDr10b suppressed NF-κB activation as well as COX-2 and iNOS expression induced by TLR3 or TLR4 agonists. CDr10b also suppressed the activation of interferon regulatory factor 3 (IRF3) and the expression of interferon inducible protein-10 (IP-10) induced by TLR3 or TLR4 agonists. These results indicate that CDr10b can modulate the TRIF-dependent pathway of TLRs and has the potential to become a new therapeutic drug for chronic inflammatory diseases.
Asunto(s)
Proteínas Adaptadoras del Transporte Vesicular/efectos de los fármacos , Compuestos de Boro/farmacología , Receptores Toll-Like/antagonistas & inhibidores , Animales , Compuestos de Boro/síntesis química , Quimiocina CXCL10/biosíntesis , Ciclooxigenasa 2/efectos de los fármacos , Factor 3 Regulador del Interferón/biosíntesis , Factor 3 Regulador del Interferón/genética , Macrófagos/efectos de los fármacos , Ratones , FN-kappa B/efectos de los fármacos , Óxido Nítrico Sintasa de Tipo II/efectos de los fármacos , Proteínas Serina-Treonina Quinasas/biosíntesis , Proteínas Serina-Treonina Quinasas/genética , Células RAW 264.7 , Transducción de Señal/efectos de los fármacos , Receptor Toll-Like 3/agonistas , Receptor Toll-Like 4/agonistasRESUMEN
When various pathogens invade a host, toll-like receptors (TLRs) play a significant role in recognizing the pathogen-associated molecular patterns carried by the pathogens to induce innate immune reaction, followed by acquired immunity reaction. TLRs have two downstream signaling pathways, the myeloid differential factor 88 (MyD88)-dependent and toll-interleukin-1 receptor domain-containing adapter inducing interferon-ß (TRIF)-dependent pathways. To evaluate the therapeutic potential of 1-[4-fluoro-2-(2-nitrovinyl)phenyl]pyrrolidine (FPP), previously synthesized in our laboratory, its effect on signal transduction via the TLR signaling pathways was examined. FPP inhibited the activation of nuclear factor-κB (NF-κB) and interferon regulatory factor 3 (IRF3) induced by TLR agonists, as well as inhibited the expression of cyclooxygenase-2, inducible nitric oxide synthase, and interferon inducible protein-10. FPP also inhibited the activation of NF-κB and IRF3 when induced by the overexpression of downstream signaling components of the TLRs. As a result, FPP has potential to become a new therapeutic drug for many inflammatory diseases.
Asunto(s)
Antiinflamatorios no Esteroideos/uso terapéutico , Inflamación/tratamiento farmacológico , Pirrolidinas/uso terapéutico , Compuestos de Vinilo/uso terapéutico , Animales , Antiinflamatorios no Esteroideos/química , Línea Celular , Quimiocina CXCL10/genética , Quimiocina CXCL10/metabolismo , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/metabolismo , Regulación hacia Abajo/efectos de los fármacos , Humanos , Factor 3 Regulador del Interferón/metabolismo , Ratones , FN-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Pirrolidinas/química , Transducción de Señal/efectos de los fármacos , Receptores Toll-Like/agonistas , Compuestos de Vinilo/químicaRESUMEN
The pathophysiological processes of inflammation can lead to a host of diseases, such as periodontitis, atherosclerosis, rheumatoid arthritis, and even cancer. The dysregulated inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) activation play important roles in the development of certain inflammatory diseases. Here, we investigated the effects of CDr10b which is originally developed for a microglia staining probe on inflammation, by modulating NF-κB activation and iNOS and COX-2 expression induced by lipopolysaccharide (LPS) in murine macrophages. The CDr10b suppressed NF-κB activation and iNOS and COX-2 expression induced by LPS. All the results suggest that CDr10b is a promising novel agent for the treatment of inflammatory diseases.
Asunto(s)
Compuestos de Boro/farmacología , Ciclooxigenasa 2/metabolismo , Lipopolisacáridos/farmacología , Óxido Nítrico Sintasa de Tipo II/metabolismo , Animales , Antiinflamatorios/farmacología , Línea Celular , Ciclooxigenasa 2/genética , Activación Enzimática/efectos de los fármacos , Expresión Génica/efectos de los fármacos , Factores Inmunológicos/farmacología , Inflamación/tratamiento farmacológico , Inflamación/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Ratones , Factor 88 de Diferenciación Mieloide/metabolismo , FN-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo II/genética , Transducción de Señal/efectos de los fármacos , Receptor Toll-Like 4/metabolismoRESUMEN
Toll-like receptors (TLRs) play an important role in the recognition of microbial pathogens and induce innate immune responses. The recognition of microbial components by TLRs triggers the activation of myeloid differential factor 88 (MyD88)- and toll-interleukin-1 receptor domain-containing adapter inducing interferon-ß (TRIF)-dependent downstream signaling pathways. Previously, we synthesized (E)-1-(2-(2-nitrovinyl)phenyl)pyrrolidine (NVPP), which contains a nitrovinyl-phenyl and pyrrolidine. To evaluate the therapeutic potential of NVPP, its effect on signal transduction via the TRIF-dependent pathway of TLRs induced by lipopolysaccharide (LPS) or polyinosinic-polycytidylic acid (poly[I:C]) was examined. NVPP inhibited LPS or poly[I:C]-induced activation of nuclear factor-κB (NF-κB) and interferon regulatory factor 3 (IRF3), and the phosphorylation of IRF3, as well as inhibiting the activation of interferon-inducible genes such as interferon inducible protein-10 (IP-10). These results suggest that NVPP can modulate TRIF-dependent signaling pathways of TLRs, potentially resulting in effective therapeutics for chronic inflammatory diseases.
Asunto(s)
Proteínas Adaptadoras del Transporte Vesicular/metabolismo , Antiinflamatorios/farmacología , Pirrolidinas/farmacología , Transducción de Señal/efectos de los fármacos , Estirenos/farmacología , Receptores Toll-Like/metabolismo , Animales , Línea Celular , Ratones , Factor 88 de Diferenciación Mieloide/metabolismoRESUMEN
AIMS: The purpose of this study was to evaluate the therapeutic potential of the helenalin in Toll-like receptor (TLR) signaling pathways. MAIN METHODS: RAW264.7 cells were transfected with a NF-κB, IFNß PRDIII-I, or IP-10 luciferase plasmid and then luciferase enzyme activities were determined by luciferase assay. The expression of iNOS, COX-2, and IP-10 and phosphorylation of IRF3 were determined by Western blotting. The levels of IP-10 were determined with culture medium by using IP-10 ELISA kit. TBK1 kinase activity was determined by MBP assay kit. KEY FINDINGS: Helenalin inhibited transcription factor NF-κB and IRF3 activation, which was induced by TLR agonists as well as its target genes, such as COX-2, iNOS, and IP-10. Helenalin attenuated ligand-independent activation of NF-κB induced by MyD88, IKKß, and p65, and IRF3 induced by TRIF, TBK1, or IRF3. Furthermore, helenalin inhibited TBK1 kinase activity in vitro. SIGNIFICANCE: TLRs are primary sensors that detect a wide variety of microbial components and play an important role in the induction of innate immune. To evaluate the therapeutic potential of helenalin, we examined its effect on signal transduction via the TLR signaling pathways. Our results suggest that beneficial effects of helenalin on chronic inflammatory diseases are mediated through modulation of TLR signaling pathways by targeting TBK1.
Asunto(s)
Proteínas Adaptadoras del Transporte Vesicular/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Sesquiterpenos/farmacología , Transducción de Señal/efectos de los fármacos , Receptores Toll-Like/metabolismo , Animales , Línea Celular , Quimiocina CXCL10/genética , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/metabolismo , Humanos , Factor 3 Regulador del Interferón/metabolismo , Lipopéptidos/farmacología , Ratones , Factor 88 de Diferenciación Mieloide/metabolismo , FN-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo II/genética , Proteínas Serina-Treonina Quinasas/genética , Sesquiterpenos de Guayano , Receptores Toll-Like/agonistasRESUMEN
Toll-like receptors (TLRs) recognize many pathogen-associated molecular patterns and induce innate immunity. TLR signaling pathways induce the activation of various transcription factors, such as nuclear factor-κB (NF-κB), leading to the induction of pro-inflammatory gene products, such as inducible nitric oxide synthase (iNOS). Here, we investigated the effect of an (E)-1-(2-(2-nitrovinyl)phenyl)pyrrolidine (NVPP), previously synthesized in our laboratory, on inflammation by modulating NF-κB activation and iNOS expression induced by TLR agonists in murine macrophages. NVPP suppressed NF-κB activation and iNOS expression induced by lipopolysaccharide (TLR4 agonist), polyriboinosinic polyribocytidylic acid (TLR3 agonist), and macrophage-activating lipopeptide 2kDa (TLR2 and TLR6 agonist). All the results suggest that NVPP is suitable for development as a new anti-inflammatory drug.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Monocitos/efectos de los fármacos , Óxido Nítrico Sintasa de Tipo II/antagonistas & inhibidores , Pirrolidinas/farmacología , Estirenos/farmacología , Receptores Toll-Like/antagonistas & inhibidores , Animales , Antiinflamatorios no Esteroideos/síntesis química , Línea Celular , Regulación de la Expresión Génica/efectos de los fármacos , Lipopéptidos/farmacología , Lipopolisacáridos/farmacología , Ratones , Monocitos/inmunología , FN-kappa B/metabolismo , Óxido Nítrico/metabolismo , Poli I-C/farmacología , Pirrolidinas/síntesis química , Estirenos/síntesis química , Receptores Toll-Like/agonistas , Activación Transcripcional/efectos de los fármacosRESUMEN
AIMS: The aim of this study was to evaluate the therapeutic potential of the phenethyl isothiocyanate (PEITC) in Toll-like receptor (TLR) signaling pathways. MAIN METHODS: To evaluate the cytotoxic nature of PEITC in RAW 264.7 cells, cytotoxicity was determined using the MTS cell viability assay. RAW264.7 cells were transfected with a nuclear factor-κB (NF-κB), interferon ß (IFNß) PRDIII-I, or interferon inducible protein-10 (IP-10) luciferase plasmid and then luciferase enzyme activities were determined by luciferase assay. The expression of inducible nitric oxide synthase (iNOS) and phosphorylation of interferon regulatory factor 3 (IRF3) were determined by Western blotting. The levels of IP-10 were determined with culture medium by using an IP-10 enzyme-linked immunosorbent assay (ELISA) kit. KEY FINDINGS: PEITC suppressed the activation of IRF3 and the expression of IP-10 induced by lipopolysaccharide (LPS) or polyinosinic-polycytidylic acid (poly[I:C]). SIGNIFICANCE: TLRs play an important role in the induction of innate immune responses for host defense against invading microbial pathogens. PEITC found in cruciferous vegetables has an effect on treatment of many chronic diseases. Our results suggest that beneficial effects of PEITC on chronic inflammatory diseases are mediated through modulation of Toll-interleukin-1 receptor domain-containing adapter inducing interferon-ß (TRIF)-dependent signaling pathway of TLRs.
Asunto(s)
Proteínas Adaptadoras del Transporte Vesicular/inmunología , Antiinflamatorios/farmacología , Anticarcinógenos/farmacología , Isotiocianatos/farmacología , Receptores Toll-Like/inmunología , Animales , Antiinflamatorios/química , Anticarcinógenos/química , Línea Celular , Inflamación/tratamiento farmacológico , Inflamación/inmunología , Factor 3 Regulador del Interferón/inmunología , Isotiocianatos/química , Lipopolisacáridos/inmunología , Ratones , Monocitos/efectos de los fármacos , Monocitos/inmunología , FN-kappa B/inmunología , Poli I-C/inmunología , Transducción de Señal/efectos de los fármacos , Verduras/químicaRESUMEN
Toll-like receptors (TLRs) play an important role by recognizing many pathogen-associated molecular patterns and inducing innate immunity. Dysregulated activation of TLR signaling pathways induces the activation of various transcription factors such as nuclear factor-κB, leading to the induction of pro-inflammatory gene products such as inducible nitric oxide synthase (iNOS). The present study investigated the effect of isobavachalcone (IBC), a natural chalcone component of Angelica keiskei, on inflammation by modulating iNOS expression induced by TLR agonists in murine macrophages. IBC suppressed iNOS expression induced by macrophage-activating lipopeptide 2-kDa, polyriboinosinic polyribocytidylic acid, or lipopolysaccharide. These results indicate the potential of IBC as a potent anti-inflammatory drug.
Asunto(s)
Antiinflamatorios/farmacología , Chalconas/farmacología , Óxido Nítrico Sintasa de Tipo II/antagonistas & inhibidores , Receptores Toll-Like/agonistas , Proteínas Adaptadoras del Transporte Vesicular/metabolismo , Animales , Línea Celular , Inhibidores Enzimáticos/farmacología , Lipopéptidos/farmacología , Lipopolisacáridos/farmacología , Ratones , Factor 88 de Diferenciación Mieloide/metabolismo , Poli I-C/farmacologíaRESUMEN
Nuclear factor-κB (NF-κB) is a transcription factor that mediates the inducible expression of a variety of genes involved in immune and inflammatory responses. NF-κB activation induces numerous proinflammatory gene products including cytokines, cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS). The divalent heavy metal mercury has been used for thousands of years. Although mercury is clearly toxic to most mammalian organ systems, especially the immune system, exposure has still increased in some areas of the world. However, the underlying toxic mechanism is not clearly identified. Here, we report biochemical evidence that mercury alone induces NF-κB activation, resulting in the induced expression of COX-2 and iNOS. The results suggest that mercury can induce inflammatory diseases by lowering host defense.