RESUMEN

Aerial parts of Artemisia asiatica (Compositae) have been traditionally used as an oriental medicine for the treatment of inflammatory and ulcerogenic diseases. In the present study, artemisolide was isolated as a nuclear factor (NF)-kappaB inhibitor from A. asiatica by activity-guided fractionation. Artemisolide inhibited NF-kappaB transcriptional activity in lipopolysaccharide (LPS)-stimulated macrophages RAW 264.7 with an IC50 value of 5.8 microM. The compound was also effective in blocking NF-kappaB transcriptional activities elicited by the expression vector encoding the NF-kappaB p65 or p50 subunits bypassing the inhibitory kB degradation signaling NF-kappaB activation. The macrophages markedly increased their PGE2 and NO production upon exposure to LPS alone. Artemisolide inhibited LPS-induced PGE2 and NO production with IC50 values of 8.7 microM and 6.4 microM, respectively, but also suppressed LPS-induced synthesis of cyclooxygenase (COX)-2 or inducible NO synthase (iNOS). Taken together, artemisolide is a NF-kappaB inhibitor that attenuates LPS-induced production of PGE2 or NO via down-regulation of COX-2 or iNOS expression in macrophages RAW 264.7. Therefore, artemisolide could represent and provide the anti-inflammatory principle associated with the traditional medicine, A. asiatica.

Asunto(s)

Antiinflamatorios/farmacología , Artemisia/química , Dinoprostona/metabolismo , Macrófagos/efectos de los fármacos , Subunidad p50 de NF-kappa B/antagonistas & inhibidores , Óxido Nítrico/metabolismo , Factor de Transcripción ReIA/antagonistas & inhibidores , Triterpenos/farmacología , Animales , Antiinflamatorios/aislamiento & purificación , Línea Celular , Ciclooxigenasa 2/metabolismo , Relación Dosis-Respuesta a Droga , Genes Reporteros , Lipopolisacáridos , Macrófagos/enzimología , Ratones , Subunidad p50 de NF-kappa B/genética , Subunidad p50 de NF-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Componentes Aéreos de las Plantas , Sesquiterpenos/farmacología , Factor de Transcripción ReIA/genética , Factor de Transcripción ReIA/metabolismo , Transfección , Triterpenos/aislamiento & purificación

RESUMEN

6-Hydroxy-7-methoxychroman-2-carboxylic acid (3-nitrophenyl)amide (CP-1158) is a synthetic chroman carboxamide with trolox-like chemical structure. In the present study, CP-1158 was found to inhibit interleukin (IL)-6 production in lipopolysaccharide (LPS)-stimulated macrophages RAW 264.7. The CP-1158 attenuated LPS-induced synthesis of IL-6 transcript but also inhibited LPS-induced IL-6 promoter activity. Further, CP-1158 attenuated LPS-induced syntheses of tumor necrosis factor (TNF)-alpha, IL-1beta, interferon-inducible protein (IP)-10 and macrophage inflammatory protein (MIP)-1beta transcripts. Nuclear factor (NF)-kappaB has been evidenced to play a major mechanism in LPS-induced expression of IL-6 or other inflammatory cytokines. CP-1158 prevented LPS-induced nuclear translocation of NF-kappaB complex and subsequently inhibited DNA binding activity of NF-kappaB complex as well as NF-kappaB transcriptional activity in macrophages RAW 264.7. However, CP-1158 did not affect LPS-induced phosphorylation and degradation of inhibitory kappaB (IkappaB). In another experiment, CP-1158 inhibited IL-6 promoter activity elicited by expression vectors encoding NF-kappaB p50 or p65 subunit. Taken together, CP-1158 inhibited LPS-induced expression of inflammatory cytokines including IL-6, targeting NF-kappaB activating pathway downstream IkappaB degradation, and thus could provide an anti-inflammatory potential of chroman carboxamide.

Asunto(s)

Antiinflamatorios/farmacología , Antioxidantes/farmacología , Cromanos/farmacología , Interleucina-6/metabolismo , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , FN-kappa B/efectos de los fármacos , FN-kappa B/metabolismo , Nitrocompuestos/farmacología , Transporte Activo de Núcleo Celular/efectos de los fármacos , Animales , Línea Celular , Quimiocina CCL4 , Quimiocina CXCL10 , Quimiocinas CXC/genética , Quimiocinas CXC/metabolismo , ADN/metabolismo , Relación Dosis-Respuesta a Droga , Vectores Genéticos , Proteínas I-kappa B/metabolismo , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Interleucina-6/genética , Proteínas Inflamatorias de Macrófagos/genética , Proteínas Inflamatorias de Macrófagos/metabolismo , Macrófagos/metabolismo , Ratones , Inhibidor NF-kappaB alfa , FN-kappa B/genética , Subunidad p50 de NF-kappa B/genética , Subunidad p50 de NF-kappa B/metabolismo , Fosforilación , Regiones Promotoras Genéticas/efectos de los fármacos , ARN Mensajero/metabolismo , Factor de Transcripción ReIA/genética , Factor de Transcripción ReIA/metabolismo , Transcripción Genética/efectos de los fármacos , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismo

RESUMEN

Furonaphthoquinone compounds have been reported to exhibit anticancer, antibacterial and antiviral properties. The molecular basis for these diverse properties is not known. 2-Methyl-2-(2-methylpropenyl)-2,3-dihydronaphthoquinone [2,3-b]furan-4,9-dione (NFD-37) is a synthetic furonaphthoquinone compound. In the present study, NFD-37 was found to inhibit interleukin (IL)-6 production in lipopolysaccharide (LPS)-stimulated murine macrophages RAW 264.7. Further, NFD-37 attenuated LPS-induced synthesis of IL-6 transcript but also inhibited LPS-induced IL-6 promoter activity. Since nuclear factor (NF)-kappaB activation has been shown to play a key role in LPS-induced IL-6 expression, the effect of NFD-37 on LPS-induced NF-kappaB activation was further analyzed. NFD-37 exhibited a dose-dependent inhibitory effect on LPS-induced phosphorylation of inhibitory kappaB alpha protein (IkappaB alpha), and subsequently inhibited LPS-induced IkappaB alpha degradation as well as NF-kappaB transcriptional activity. In another experiment, NFD-37 inhibited both IL-6 promoter activity and NF-kappaB transcriptional activity elicited by an expression vector encoding IkappaB kinase beta. Taken together, NFD-37 down-regulated LPS-induced IL-6 expression through NF-kappaB activation, which could provide a pharmacological basis for the anti-inflammatory properties of furonaphthoquinone analogs.

Asunto(s)

Furanos/farmacología , Interleucina-6/metabolismo , Macrófagos/efectos de los fármacos , Naftoquinonas/farmacología , Animales , Línea Celular , Relación Dosis-Respuesta a Droga , Expresión Génica/efectos de los fármacos , Quinasa I-kappa B/genética , Quinasa I-kappa B/metabolismo , Proteínas I-kappa B/genética , Proteínas I-kappa B/metabolismo , Interleucina-6/genética , Lipopolisacáridos/farmacología , Macrófagos/citología , Macrófagos/metabolismo , Inhibidor NF-kappaB alfa , FN-kappa B/genética , FN-kappa B/metabolismo , Subunidad p50 de NF-kappa B/genética , Subunidad p50 de NF-kappa B/metabolismo , Fosforilación/efectos de los fármacos , Regiones Promotoras Genéticas/efectos de los fármacos , Factor de Transcripción ReIA/genética , Factor de Transcripción ReIA/metabolismo , Transfección

RESUMEN

Tyrosinase is a key enzyme for melanin biosynthesis, and hyperpigmentation disorders are associated with abnormal accumulation of melanin pigments, which can be improved by treatment with depigmenting agents. In the present study, piperlonguminine from Piper longum was discovered to inhibit melanin production in melanoma B16 cells stimulated with alpha-melanocyte stimulating hormone (alpha-MSH), 3-isobutyl-1-methylxanthine or protoporphyrin IX, where the compound exhibited stronger depigmenting efficacy than kojic acid. However, piperlonguminine did not affect 1-oleoyl-2-acetyl-sn-glycerol-induced melanogenesis and did not affect protein kinase C-mediated melanin production. Surprisingly, piperlonguminine did not inhibit the catalytic activity of cell-free tyrosinase from melanoma B16 cells but rather suppressed tyrosinase mRNA expression. This effect was attributed to the inhibitory action of piperlonguminine on alpha-MSH-induced signaling through cAMP to the cAMP responsive element binding protein that in turn regulates the expression of the microphthalmia-associated transcription factor, a key activator of the tyrosinase promoter. This study demonstrates that piperlonguminine is an efficient depigmenting agent with a novel mechanism of action.

Asunto(s)

Dioxolanos/metabolismo , Melaninas/biosíntesis , Monofenol Monooxigenasa/metabolismo , Piper/química , Extractos Vegetales/metabolismo , 1-Metil-3-Isobutilxantina/metabolismo , Animales , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Diglicéridos/metabolismo , Dioxolanos/química , Dioxolanos/farmacología , Regulación hacia Abajo , Humanos , Melanoma , Factor de Transcripción Asociado a Microftalmía/metabolismo , Estructura Molecular , Monofenol Monooxigenasa/genética , Inhibidores de Fosfodiesterasa/metabolismo , Extractos Vegetales/química , Extractos Vegetales/farmacología , Protoporfirinas/metabolismo , ARN Mensajero/metabolismo , alfa-MSH/metabolismo

RESUMEN

2-Methyl-2-(2-methylpropenyl)-2,3-dihydronaphthoquinone[2,3-b]furan-4,9-dione (NFD-37) is a synthetic furonaphthoquinone compound. In this study, we determined that NFD-37 could inhibit the lipopolysaccharide (LPS)-induced production of inflammatory mediators in macrophages RAW 264.7. This compound inhibited LPS-induced nitric oxide (NO) or prostaglandin (PG) E2 production in dose-dependent manners, with IC50 values of 7.2 microM and 5.3 microM, respectively. As the positive controls, pyrrolidine dithiocarbamate (30 microM) exhibited a 57% inhibition of NO production, and NS-398 (1 microM) manifested a 48% inhibition of PGE2 production. The inhibitory effects of NFD-37 on NO and PGE2 production were determined to occur in conjunction with the suppression of inducible NO synthase or cyclooxygenase-2 expression. NFD-37 also inhibited the production of LPS-inducible tumor necrosis factor-alpha, interleukin (IL)-1beta and IL-6, at IC50 values of 4.8-8.9 microM. We also determined the anti-inflammatory efficacy of NFD-37 using carrageenin-induced paw edema in experimental mice.

Asunto(s)

Furanos/farmacología , Mediadores de Inflamación/metabolismo , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Naftoquinonas/farmacología , Animales , Western Blotting , Línea Celular , Supervivencia Celular/efectos de los fármacos , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/metabolismo , Dinoprostona/metabolismo , Relación Dosis-Respuesta a Droga , Furanos/química , Interleucina-1/metabolismo , Interleucina-6/metabolismo , Macrófagos/citología , Macrófagos/metabolismo , Estructura Molecular , Naftoquinonas/química , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transcripción Genética/efectos de los fármacos , Factor de Necrosis Tumoral alfa/metabolismo

RESUMEN

2-Methyl-2-(2-methylpropenyl)-2,3-dihydronaphtho[2,3-b]furan-4,9-dione (NFD-37) is a synthetic furonaphthoquinone compound. In the present study, the NFD-37 compound was found to inhibit prostaglandin (PG) E(2) production in lipopolysaccharide (LPS)-stimulated macrophages RAW 264.7. NFD-37 compound exhibited a preferred inhibition on enzyme activity of cyclooxygenase (COX)-2 over COX-1. Further, NFD-37 compound attenuated LPS-induced synthesis of both mRNA and protein of COX-2, and suppressed LPS-induced COX-2 promoter activity in the macrophages, indicating that the furonaphthoquinone compound could down-regulate LPS-induced COX-2 expression at the transcription level. Even though COX-2 promoter behaves as a sophisticated biosensor for host defense, nuclear factor (NF)-kappaB activation has been evidenced to play a major mechanism for LPS-induced COX-2 expression in macrophages. NFD-37 compound exhibited a dose-dependent inhibitory effect on LPS-induced phosphorylation of inhibitory kappaBalpha (IkappaBalpha) protein, and subsequently inhibited IkappaBalpha degradation, DNA binding activity of NF-kappaB complex as well as NF-kappaB transcriptional activity in macrophages RAW 264.7. In another experiment, NFD-37 compound inhibited both COX-2 promoter activity and GST-IkappaBalpha phosphorylation elicited by an expression vector encoding IkappaB kinase beta. Taken together, NFD-37 compound inhibited enzyme activity of COX-2 but also suppressed COX-2 expression depending on NF-kappaB activation, and thus could provide an invaluable tool to investigate pharmacological potential in the excess PG-related disorders.

Asunto(s)

Inhibidores de la Ciclooxigenasa/farmacología , Furanos/farmacología , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Naftoquinonas/farmacología , Prostaglandina-Endoperóxido Sintasas/metabolismo , Animales , Secuencia de Bases , Línea Celular , Ciclooxigenasa 2 , Inhibidores de la Ciclooxigenasa 2 , Cartilla de ADN , Dinoprostona/biosíntesis , Regulación hacia Abajo/efectos de los fármacos , Macrófagos/enzimología , Macrófagos/metabolismo , Ratones , FN-kappa B/metabolismo , Regiones Promotoras Genéticas , Prostaglandina-Endoperóxido Sintasas/genética , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transcripción Genética

RESUMEN

Quercetin 3-O-beta-(2''-galloyl)-rhamnopyranoside (QGR) is a naturally occurring quercitrin gallate, a polyphenolic compound isolated from Persicaria lapathifolia (Polygonaceae). In the present study, QGR compound was discovered to have inhibitory effect on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated macrophages RAW 264.7. QGR compound attenuated LPS-induced synthesis of both mRNA and protein of inducible nitric oxide synthase (iNOS), in parallel, and inhibited LPS-induced luciferase expression as a reporter of iNOS promoter activity in the macrophages. As a mechanism of the anti-inflammatory action shown by QGR compound, suppression of nuclear factor (NF)-kappaB activation has been documented. QGR compound exhibited inhibitory effect on LPS-mediated NF-kappaB transcriptional activity in macrophages RAW 264.7. Furthermore, the compound inhibited LPS-mediated nuclear translocation of NF-kappaB p65 and DNA binding activity of NF-kappaB complex, in parallel, but did not influence LPS-mediated IkappaBalpha degradation. Taken together, QGR compound suppressed LPS-mediated NF-kappaB activation, specifically to nuclear localization step of NF-kappaB p65, which was attributable to its down-regulatory action on LPS-induced NO production and iNOS expression.

Asunto(s)

Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Activación de Macrófagos/fisiología , FN-kappa B/fisiología , Óxido Nítrico Sintasa/biosíntesis , Quercetina/análogos & derivados , Quercetina/farmacología , Animales , Línea Celular , Relación Dosis-Respuesta a Droga , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/fisiología , Regulación Enzimológica de la Expresión Génica/fisiología , Lipopolisacáridos/farmacología , Activación de Macrófagos/efectos de los fármacos , Ratones , Óxido Nítrico Sintasa/genética , Óxido Nítrico Sintasa de Tipo II , Quercetina/química

RESUMEN

Toll-like receptor 4 (TLR4) is known to play an important role in innate immune responses. In the present study, chemically synthetic compound of N(1)-benzyl-4-methylbenzene-1,2-diamine (BMD) was discovered to inhibit nitric oxide (NO) production in macrophages RAW 264.7 stimulated with lipopolysaccharide (LPS) or fibronectin as TLR4 activators. The BMD compound attenuated LPS-induced synthesis of both mRNA and protein of NO synthase (iNOS), and inhibited LPS or fibronectin-mediated iNOS promoter activity, indicating that the compound down-regulated iNOS expression at transcription level. As a mode of the anti-inflammatory action shown by BMD compound, inhibitory effect on nuclear factor (NF)-kappaB activation was also investigated in macrophages RAW 264.7 stimulated with the TLR4 activators.

Asunto(s)

Antiinflamatorios no Esteroideos/farmacología , Benceno/farmacología , Diaminas/farmacología , Receptor Toll-Like 4/biosíntesis , Animales , Antiinflamatorios no Esteroideos/química , Benceno/química , Línea Celular , Diaminas/química , Relación Dosis-Respuesta a Droga , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Regulación Enzimológica de la Expresión Génica , Ratones , Receptor Toll-Like 4/antagonistas & inhibidores

RESUMEN

6-Hydroxy-7-methoxychroman-2-carboxylic acid phenylamide (KL-1156) is a novel chemically synthetic compound. In the present study, the chroman KL-1156 compound was found to inhibit lipopolysaccharide (LPS)-induced nitric oxide production in macrophages RAW 264.7. KL-1156 compound attenuated LPS-induced synthesis of both mRNA and protein of inducible nitric oxide synthase (iNOS), in parallel, and inhibited LPS-induced iNOS promoter activity, indicating that the chroman compound down-regulated iNOS expression at transcription level. As a mechanism of the anti-inflammatory action shown by KL-1156 compound, suppression of nuclear factor (NF)-kappaB has been documented. KL-1156 compound exhibited a dose-dependent inhibitory effect on LPS-induced NF-kappaB transcriptional activity in macrophages RAW 264.7. Furthermore, the compound inhibited LPS-induced nuclear translocation of NF-kappaB p65 and DNA binding activity of NF-kappaB complex, in parallel, but did not affect IkappaBalpha degradation. Taken together, this study demonstrated that chroman KL-1156 compound interfered with nuclear translocation step of NF-kappaB p65, which was attributable to its anti-inflammatory action.

Asunto(s)

Cromanos/farmacología , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , FN-kappa B/metabolismo , Óxido Nítrico/metabolismo , Transporte Activo de Núcleo Celular/fisiología , Animales , Línea Celular , Cromanos/química , ADN/metabolismo , Regulación Enzimológica de la Expresión Génica , Proteínas I-kappa B/metabolismo , Macrófagos/citología , Macrófagos/inmunología , Ratones , Estructura Molecular , Inhibidor NF-kappaB alfa , FN-kappa B/inmunología , Óxido Nítrico Sintasa/genética , Óxido Nítrico Sintasa/metabolismo , Óxido Nítrico Sintasa de Tipo II , Regiones Promotoras Genéticas , Factor de Transcripción ReIA , Transcripción Genética

RESUMEN

N1-Benzyl-4-methylbenzene-1,2-diamine (JSH-21) and its analogs were chemically synthesized and their anti-inflammatory potentials investigated. JSH-21 inhibited nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated macrophages RAW 264.7 in a dose-dependent manner, with an IC50 value of 9.2 microM, where pyrrolidine dithiocarbamate and parthenolide as positive controls exhibited IC50 values of 29.3 and 3.6 microM, respectively. The inhibitory effect of JSH-21 on the NO production was attributable to its down-regulatory action on LPS-inducible NO synthase (iNOS), which was documented by iNOS promoter activity. In the mechanism of the anti-inflammatory action, JSH-21 exhibited inhibitory effects on LPS-induced DNA binding activity and transcriptional activity of nuclear factor-kappa B (NF-kappaB). Structural analogs of JSH-21 also inhibited both the LPS-induced NO production and NF-kappaB transcriptional activity, where diamine substitution at positions 1 and 2 of JSH-21 seems to play an important role in the anti-inflammatory activity.

Asunto(s)

Aminas/farmacología , Antiinflamatorios no Esteroideos/farmacología , Compuestos de Bencilo/farmacología , Lipopolisacáridos/farmacología , Macrófagos/metabolismo , FN-kappa B/metabolismo , Óxido Nítrico/biosíntesis , Transcripción Genética/efectos de los fármacos , Línea Celular , ADN/efectos de los fármacos , ADN/metabolismo , Relación Dosis-Respuesta a Droga , Ensayo de Cambio de Movilidad Electroforética , Humanos , Macrófagos/efectos de los fármacos , Espectroscopía de Resonancia Magnética , FN-kappa B/genética , Óxido Nítrico Sintasa/antagonistas & inhibidores , Óxido Nítrico Sintasa de Tipo II

RESUMEN

Nuclear factor (NF)-cB is a transcription factor regulating the expression of inflammatory and immune genes. In the present study, an extract from stem bark of Cinnamomum cassia Blume(Lauraceae) was discovered to have an inhibitory effect on LPS-induced NF-KB transcriptional activity, which was determined using macrophages RAW 264.7 transfected stably with an alkaline phosphatase reporter construct containing four copies of the NF-KB binding KB sequence. Following activity-guided fractionation, trans-cinnamaldehyde and 2-methoxycinnamaldehyde were identified as the NF-KB inhibitors from C cassia with IC50 values of 43 MM and 31 pM, respectively. As a positive control, caffeic acid phenethyl ester (CAPE) showed an IC50 value of 2 uM on NF-KB transcriptional activity. Both trans-cinnamaldehyde and 2-methoxycinnamaldehyde inhibited LPS-induced DNA binding activity of NF-KB in addition to NF-KB transcription-al activity.

Asunto(s)

Acroleína/análogos & derivados , Acroleína/farmacología , Cinnamomum aromaticum , Macrófagos/metabolismo , FN-kappa B/antagonistas & inhibidores , Fitoterapia , Acroleína/administración & dosificación , Acroleína/uso terapéutico , Animales , Concentración 50 Inhibidora , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Ratones , FN-kappa B/biosíntesis , Extractos Vegetales/administración & dosificación , Extractos Vegetales/farmacología , Extractos Vegetales/uso terapéutico , Tallos de la Planta

RESUMEN

4-Methyl-N1-(3-phenyl-propyl)-benzene-1,2-diamine (JSH-23) is a novel chemically synthetic compound. The aromatic diamine JSH-23 compound exhibited inhibitory effect with an IC(50) value of 7.1 microM on nuclear factor (NF)-kappaB transcriptional activity in lipopolysaccharide (LPS)-stimulated macrophages RAW 264.7, and interfered LPS-induced nuclear translocation of NF-kappaB without affecting IkappaB degradation. This mechanism of action is very rare for controlling NF-kappaB activation. Furthermore, the compound inhibited not only LPS-induced expressions of tumor necrosis factor-alpha, interleukin (IL)-1beta, IL-6 and inducible nitric oxide synthase and cyclooxygenase-2 but also LPS-induced apoptosis of the RAW 264.7 cells.

Asunto(s)

Núcleo Celular/metabolismo , Proteínas I-kappa B/metabolismo , Lipopolisacáridos/farmacología , Macrófagos/metabolismo , FN-kappa B/metabolismo , Animales , Línea Celular , Núcleo Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Citocinas/genética , Escherichia coli , Regulación de la Expresión Génica/efectos de los fármacos , Inflamación , Cinética , Ratones , Inhibidor NF-kappaB alfa , Óxido Nítrico Sintasa/genética , Óxido Nítrico Sintasa de Tipo II , Transporte de Proteínas/efectos de los fármacos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa

RESUMEN

2-Hydroxymuconic semialdehyde (2-HMS) dehydrogenase catalyzes the conversion of 2-HMS to 4-oxalocrotonate, which is a step in the meta cleavage pathway of aromatic hydrocarbons in bacteria. A tomC gene that encodes 2-HMS dehydrogenase of Burkholderia cepacia G4, a soil bacterium that can grow on toluene, cresol, phenol, or benzene, was overexpressed into E. coli HB101, and its gene product was characterized in this study. 2-HMS dehydrogenase from B. cepacia G4 has a high catalytic efficiency in terms of Vmax/Km towards 2-hydroxy-5-methylmuconic semialdehyde followed by 2-HMS but has a very low efficiency for 5-chloro-2-hydroxymuconic semialdehyde. However, the enzyme did not utilize 2-hydroxy-6-oxo-hepta-2,4-dienoic acid and 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid as substrates. The molecular weight of 2-HMS dehydrogenase from B. cepacia G4 was predicted to be 52 kDa containing 485 amino acid residues from the nucleotide sequence of the tomC gene, and it exhibited the highest identity of 78% with the amino acid sequence of 2-HMS dehydrogenase that is encoded in the aphC gene of Comamonas testosteroni TA441. 2-HMS dehydrogenase from B. cepacia G4 showed a significant phylogenetic relationship not only with other 2-HMS dehydrogenases, but also with different dehydrogenases from evolutionarily distant organisms.

Asunto(s)

Aldehído Oxidorreductasas/genética , Burkholderia cepacia/enzimología , Burkholderia cepacia/genética , Filogenia , Plásmidos/genética , Plásmidos/metabolismo

RESUMEN

N(1)-Benzyl-4-methylbenzene-1,2-diamine (JSH-21) and its analogs were chemically synthesized and their anti-inflammatory potentials investigated. JSH-21 inhibited nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated macrophages RAW 264.7 in a dose-dependent manner, with an IC50 value of 9.2 µM, where pyrrolidine dithiocarbamate and parthenolide as positive controls exhibited IC50 values of 29.3 and 3.6 µM, respectively. The inhibitory effect of JSH-21 on the NO production was attributable to its down-regulatory action on LPS-induc-ible NO synthase (iNOS), which was documented by iNOS promoter activity. In the mechanism of the anti-inflammatory action, JSH-21 exhibited inhibitory effects on LPS-induced DNA binding activity and transcriptional activity of nuclear factor-kappa B(NF-KB). Structural analogs of JSH-21 also inhibited both the LPS-induced NO production andNF-KB transcriptional activity, where diamine substitution at positions 1 and 2 of JSH-21 seems to play an important role in the anti-inflammatory activity.

RESUMEN

Skin hyperpigmentations such as melasma, freckles and senile lentigines can be subjectively treated by depigmenting agents. In our ongoing study to find melanogenesis inhibitors from natural sources, Piper longum L (fruits, Piperaceae) was discovered to have an inhibitory effect on alpha-melanocyte-stimulating hormone (alpha-MSH)-induced melanogenesis in melanoma B16 cells. Piperlonguminine has been identified as the melanogenesis inhibitor from P. longum by activity-guided extraction and isolation. The compound showed dose-dependent inhibitory effects with 85.1 +/- 4.9% inhibition at 25 microM, 62.1 +/- 6.1% at 12.5 microM, 36.4 +/- 4.6% at 6.3 microM and 18.4 +/- 5.1% at 3.1 microM on alpha-MSH-induced melanogenesis, showing an IC50 value of 9.6 microM. As a positive control, kojic acid exhibited an IC50 value of 44.6 microM on the melanogenesis. As to the mode of action, piperlonguminine showed an inhibitory effect on alpha-MSH-induced tyrosinase synthesis, documented by Western immunoblot analysis. However, piperlonguminine did not show an inhibitory effect on tyrosinase activity or a direct depigmenting effect of melanin.

Asunto(s)

Dioxolanos/farmacología , Fitoterapia , Piper , Extractos Vegetales/farmacología , alfa-MSH/antagonistas & inhibidores , alfa-MSH/efectos de los fármacos , Animales , Línea Celular Tumoral/efectos de los fármacos , Dioxolanos/administración & dosificación , Dioxolanos/uso terapéutico , Relación Dosis-Respuesta a Droga , Frutas , Melaninas/biosíntesis , Ratones , Monofenol Monooxigenasa/biosíntesis , Trastornos de la Pigmentación/tratamiento farmacológico , Extractos Vegetales/administración & dosificación , Extractos Vegetales/uso terapéutico

RESUMEN

The anti-inflammatory activity of alpha-viniferin, a trimer of resveratrol, has been demonstrated in an animal model of carrageenin-induced paw edema, and inhibitory effects of the compound on cyclooxygenase (COX) and inducible nitric oxide synthase (iNOS) have been investigated in order to understand the mode of the observed action. alpha-Viniferin at doses > 30 mg/kg ( p. o.) or > 3 mg/kg ( i. v.) showed significant anti-inflammatory activity on carrageenin-induced paw edema in mice. alpha-Viniferin showed an inhibitory effect with an IC (50) value of 4.9 microM on COX-2 activity but a very weak inhibitory effect with 55.2 +/- 2.1 % of the control (100 %) at 100 microM on COX-1 activity. alpha-Viniferin at doses of 3 microM to 10 microM inhibited the synthesis of COX-2 transcript in lipopolysaccharide (LPS)-activated murine macrophages Raw264.7. alpha-Viniferin showed an IC50 value of 2.7 microM on nitric oxide (NO) production in LPS-activated Raw264.7 cells when alpha-viniferin and LPS were treated simultaneously, but did not inhibit the NO production when alpha-viniferin was treated at 12 h after LPS stimulation. alpha-Viniferin inhibited synthesis of iNOS transcript with an IC50 value of 4.7 microM. Consequently, the inhibitory effect of alpha-viniferin on the release of prostanoids and NO could play an important role to show anti-inflammatory action.

Asunto(s)

Antiinflamatorios no Esteroideos/farmacología , Benzofuranos/farmacología , Caragana , Edema/prevención & control , Isoenzimas/antagonistas & inhibidores , Óxido Nítrico Sintasa/antagonistas & inhibidores , Fitoterapia , Administración Oral , Animales , Antiinflamatorios no Esteroideos/administración & dosificación , Antiinflamatorios no Esteroideos/uso terapéutico , Benzofuranos/administración & dosificación , Benzofuranos/uso terapéutico , Carragenina , Línea Celular , Ciclooxigenasa 2 , Inhibidores de la Ciclooxigenasa 2 , Inhibidores de la Ciclooxigenasa/administración & dosificación , Inhibidores de la Ciclooxigenasa/farmacología , Inhibidores de la Ciclooxigenasa/uso terapéutico , Cartilla de ADN , Relación Dosis-Respuesta a Droga , Edema/inducido químicamente , Concentración 50 Inhibidora , Inyecciones Intravenosas , Macrófagos/efectos de los fármacos , Macrófagos/enzimología , Masculino , Ratones , Ratones Endogámicos ICR , Óxido Nítrico Sintasa de Tipo II , Prostaglandina-Endoperóxido Sintasas , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Estilbenos/administración & dosificación , Estilbenos/farmacología , Estilbenos/uso terapéutico

RESUMEN

Soy is a main source of isoflavonoids which are of high dietary intake for the oriental population. In this study, the anti-inflammatory action of sophoricoside, an isoflavone glycoside isolated from immature fruits of Sophora japonica L. (Leguminosae), has been demonstrated. When administered orally at > 100 mg/kg or injected intravenously at > 10 mg/kg, sophoricoside showed significant reduction of carrageenin-induced paw edema in mice. Sophoricoside has been identified as a selective inhibitor of cyclooxygenase (COX)-2 activity with an IC50 value of 3.3 microM.

Asunto(s)

Antiinflamatorios no Esteroideos/farmacología , Benzopiranos/farmacología , Inhibidores de la Ciclooxigenasa/farmacología , Isoflavonas/farmacología , Fitoterapia , Sophora , Administración Oral , Animales , Antiinflamatorios no Esteroideos/administración & dosificación , Antiinflamatorios no Esteroideos/uso terapéutico , Benzopiranos/administración & dosificación , Benzopiranos/uso terapéutico , Carragenina , Ciclooxigenasa 2 , Inhibidores de la Ciclooxigenasa 2 , Inhibidores de la Ciclooxigenasa/administración & dosificación , Inhibidores de la Ciclooxigenasa/uso terapéutico , Relación Dosis-Respuesta a Droga , Edema/inducido químicamente , Edema/prevención & control , Concentración 50 Inhibidora , Inyecciones Intravenosas , Isoenzimas/antagonistas & inhibidores , Isoenzimas/efectos de los fármacos , Isoflavonas/administración & dosificación , Isoflavonas/uso terapéutico , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Ratones , Prostaglandina-Endoperóxido Sintasas/efectos de los fármacos

RESUMEN

Soy, high dietary intake for the oriental population, is a main source of isoflavonoids. Sophoricoside (SOP) an isoflavone glycoside was isolated from immature fruits of Sophora japonica (Leguminosae family) and its inhibitory effect on chemical mediators involved in inflammatory response was investigated in this study. SOP inhibited the interleukin (IL)-6 bioactivity with an IC50 value of 6.1 microM whereas it had no effects on IL-1beta and TNF-alpha bioactivities. SOP was identified as a selective inhibitor of cyclooxygenase (COX)-2 activity with an IC50 value of 4.4 microM, but did not show inhibitory effect on the synthesis of COX-2. However, SOP had no effect on the production of reactive oxygen species including superoxide anions and nitric oxide. These results revealed that in vitro anti-inflammatory action of SOP is significantly different from that of genistein known as a phytoestrogen of soy products. This experimental study has documented an importance of dietary soy isoflavonoids as multifunctional agents beneficial to human health, and will help to clarify protective mechanisms of SOP against inflammatory conditions.

Asunto(s)

Antiinflamatorios/farmacología , Benzopiranos/farmacología , Inflamación/tratamiento farmacológico , Inflamación/prevención & control , Interleucina-6/antagonistas & inhibidores , Isoenzimas/antagonistas & inhibidores , Óxido Nítrico/antagonistas & inhibidores , Especies Reactivas de Oxígeno/antagonistas & inhibidores , Animales , Ciclooxigenasa 1 , Ciclooxigenasa 2 , Humanos , Técnicas In Vitro , Inflamación/inducido químicamente , Interleucina-1/metabolismo , Isoenzimas/efectos adversos , Corea (Geográfico) , Proteínas de la Membrana , Ratones , Óxido Nítrico/metabolismo , Estructuras de las Plantas/química , Prostaglandina-Endoperóxido Sintasas/efectos adversos , Especies Reactivas de Oxígeno/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo

RESUMEN

Tyrosinase is responsible for the molting process in insects, undesirable browning of fruits and vegetables, and coloring of skin, hair, and eyes in animals. To clarify the mechanism of the depigmenting property of hydroxystilbene compounds, inhibitory actions of oxyresveratrol and its analogs on tyrosinases from mushroom and murine melanoma B-16 have been elucidated in this study. Oxyresveratrol showed potent inhibitory effect with an IC(50) value of 1.2 microm on mushroom tyrosinase activity, which was 32-fold stronger inhibition than kojic acid, a depigmenting agent used as the cosmetic material with skin-whitening effect and the medical agent for hyperpigmentation disorders. Hydroxystilbene compounds of resveratrol, 3,5-dihydroxy-4'-methoxystilbene, and rhapontigenin also showed more than 50% inhibition at 100 microm on mushroom tyrosinase activity, but other methylated or glycosylated hydroxystilbenes of 3,4'-dimethoxy-5-hydroxystilbene, trimethylresveratrol, piceid, and rhaponticin did not inhibit significantly. None of the hydroxystilbene compounds except oxyresveratrol exhibited more than 50% inhibition at 100 microm on l-tyrosine oxidation by murine tyrosinase activity; oxyresveratrol showed an IC(50) value of 52.7 microm on the enzyme activity. The kinetics and mechanism for inhibition of mushroom tyrosinase exhibited the reversibility of oxyresveratrol as a noncompetitive inhibitor with l-tyrosine as the substrate. The interaction between oxyresveratrol and tyrosinase exhibited a high affinity reflected in a K(i) value of 3.2-4.2 x 10(-7) m. Oxyresveratrol did not affect the promoter activity of the tyrosinase gene in murine melanoma B-16 at 10 and 100 microm. Therefore, the depigmenting effect of oxyresveratrol works through reversible inhibition of tyrosinase activity rather than suppression of the expression and synthesis of the enzyme. The number and position of hydroxy substituents seem to play an important role in the inhibitory effects of hydroxystilbene compounds on tyrosinase activity.

Asunto(s)

Inhibidores Enzimáticos/farmacología , Monofenol Monooxigenasa/antagonistas & inhibidores , Monofenol Monooxigenasa/genética , Extractos Vegetales/farmacología , Estilbenos/farmacología , Agaricales/enzimología , Animales , Antioxidantes/farmacología , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Cinética , Melanoma Experimental , Ratones , Regiones Promotoras Genéticas/efectos de los fármacos , Pironas/farmacología , Relación Estructura-Actividad , Células Tumorales Cultivadas