RESUMO
Cisplatin (cis-diaminedichloroplatinum-II) is an extensively used chemotherapeutic agent, and one of its most adverse effects is ototoxicity. A number of studies have demonstrated that these effects are related to oxidative stress and DNA damage. However, the precise mechanism underlying cisplatin-associated ototoxicity is still unclear. The cofactor nicotinamide adenine dinucleotide (NAD(+)) has emerged as a key regulator of cellular energy metabolism and homeostasis. Here, we demonstrate for the first time that, in cisplatin-mediated ototoxicity, the levels and activities of SIRT1 are suppressed by the reduction of intracellular NAD(+) levels. We provide evidence that the decrease in SIRT1 activity and expression facilitated by increasing poly(ADP-ribose) transferase (PARP)-1 activation and microRNA-34a through p53 activation aggravates cisplatin-mediated ototoxicity. Moreover, we show that the induction of cellular NAD(+) levels using ß-lapachone (ß-Lap), whose intracellular target is NQO1, prevents the toxic effects of cisplatin through the regulation of PARP-1 and SIRT1 activity. These results suggest that direct modulation of cellular NAD(+) levels by pharmacological agents could be a promising therapeutic approach for protection from cisplatin-induced ototoxicity.
Assuntos
Antineoplásicos/efeitos adversos , Cisplatino/efeitos adversos , Perda Auditiva/induzido quimicamente , NAD(P)H Desidrogenase (Quinona)/metabolismo , NAD/metabolismo , Animais , Antineoplásicos/farmacologia , Cisplatino/farmacologia , Perda Auditiva/genética , Perda Auditiva/metabolismo , Perda Auditiva/patologia , Masculino , Camundongos , Camundongos Knockout , MicroRNAs/genética , MicroRNAs/metabolismo , NAD/genética , NAD(P)H Desidrogenase (Quinona)/genética , Poli(ADP-Ribose) Polimerase-1 , Poli(ADP-Ribose) Polimerases/genética , Poli(ADP-Ribose) Polimerases/metabolismo , Sirtuína 1/genética , Sirtuína 1/metabolismo , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismoRESUMO
We studied the effect of mokko lactone (ML) isolated from the roots of Saussurea lappa (Compositae), a plant that is used for medicinal purposes in Korea, on the induction of apoptosis in human leukemia HL-60 cells. ML was cytotoxic to HL-60 cells, and this cytotoxic effect of ML appears to be attributable to its induction of apoptotic cell death, as ML induced nuclear morphologic changes and internucleosomal DNA fragmentation and increased the proportion of Annexin V-positive cells and the activity of caspase-3. Further studies revealed that the induction of apoptosis by ML was associated with the loss of mitochondrial membrane potential. Collectively, our results suggest that apoptosis induced by ML in HL-60 cells was executed by a collapse of mitochondrial membrane potential followed by the activation of caspase-3. This is the first report on the mechanism of apoptosis-inducing effect of ML.
Assuntos
4-Butirolactona/análogos & derivados , 4-Butirolactona/farmacologia , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Células HL-60/efeitos dos fármacos , Membranas Intracelulares/efeitos dos fármacos , Lactonas/farmacologia , Mitocôndrias/efeitos dos fármacos , Saussurea , Sesquiterpenos/farmacologia , 4-Butirolactona/isolamento & purificação , Caspase 3 , Caspases/análise , Caspases/metabolismo , Fragmentação do DNA , Humanos , Membranas Intracelulares/patologia , Coreia (Geográfico) , Lactonas/isolamento & purificação , Potenciais da Membrana/efeitos dos fármacos , Mitocôndrias/patologia , Raízes de Plantas/química , Saussurea/química , Sesquiterpenos/isolamento & purificaçãoRESUMO
Sesquiterpene lactones have raised considerable interest because of their ability to block the activation of nuclear transcription factor-kappaB (NF-kappaB). NF-kappaB plays an important role in the resistance of cancer cells to the induction of apoptosis by anticancer drugs and tumor necrosis factor-alpha (TNF-alpha). Pharmacological inhibition of NF-kappaB offers the promise of enhancing the efficacy of anticancer therapies. Here, we demonstrate that dehydrocostus lactone (DL), the major sesquiterpene lactone isolated from the roots of Saussurea lappa, inhibits NF-kappaB activation by preventing TNF-alpha-induced degradation and phosphorylation of its inhibitory protein I-kappaB alpha in human leukemia HL-60 cells and that DL renders HL-60 cells susceptible to TNF-alpha-induced apoptosis by enhancing caspase-8 and caspase-3 activities.
Assuntos
Apoptose/efeitos dos fármacos , Lactonas/administração & dosagem , Sesquiterpenos/administração & dosagem , Fator de Necrose Tumoral alfa/administração & dosagem , Antineoplásicos/administração & dosagem , Antineoplásicos/isolamento & purificação , Caspase 3 , Caspase 8 , Caspases/metabolismo , Sinergismo Farmacológico , Células HL-60 , Humanos , Lactonas/isolamento & purificação , NF-kappa B/metabolismo , Proteínas de Neoplasias/biossíntese , Fitoterapia , Saussurea/química , Sesquiterpenos/isolamento & purificaçãoRESUMO
We investigated the effects of the gallotannin penta-O-galloyl-beta-d-glucose (PGG) on interleukin (IL)-8 gene expression and nuclear factor (NF)-kappaB activation. PGG inhibited IL-8 production and gene expression in human monocytic U937 cells stimulated with phorbol myristate acetate (PMA), as measured by enzyme-linked immunosorbent assay and reverse transcription-polymerase chain reaction analysis, respectively. PGG also inhibited PMA-mediated NF-kappaB activation, as measured by electromobility shift assay. Furthermore, PGG prevented PMA-mediated degradation of the NF-kappaB inhibitory protein I-kappaBalpha, as measured by Western blot analysis. PGG also inhibited both IL-8 production and NF-kappaB activation in the U937 cells stimulated with tumor necrosis factor-alpha. These results suggest that PGG, a major constituent of the root cortex of Paeonia suffruticosa ANDREWS, can inhibit IL-8 gene expression by a mechanism involving its inhibition of NF-kappaB activation, which is dependent on I-kappaBalpha degradation.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Taninos Hidrolisáveis/farmacologia , Interleucina-8/antagonistas & inibidores , NF-kappa B/antagonistas & inibidores , Acetato de Tetradecanoilforbol/farmacologia , Transporte Ativo do Núcleo Celular , Western Blotting , Ensaio de Desvio de Mobilidade Eletroforética , Ensaio de Imunoadsorção Enzimática , Regulação da Expressão Gênica , Humanos , Proteínas I-kappa B/metabolismo , Interleucina-8/biossíntese , Interleucina-8/genética , Transporte Proteico , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , Células U937RESUMO
Ginsenosides from Panax ginseng are metabolized by human intestinal bacteria after oral administration of ginseng extract. 20(S)-Protopanaxatriol (PPT) is one of the major metabolites of ginsenosides. Inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) are important enzymes that mediate inflammatory processes. Improper up-regulation of iNOS and/or COX-2 has been associated with the pathogenesis of inflammatory diseases and certain types of human cancers. Here, we investigated whether PPT could modulate iNOS and COX-2 expressions in RAW 264.7 macrophages stimulated with the endotoxin lipopolysaccharide (LPS). We found that PPT blocked the increase in LPS-induced iNOS and COX-2 expressions through inactivation of nuclear factor-kappaB by preventing I-kappaBalpha phosphorylation and degradation. Thus, it may be possible to develop PPT as a useful agent for chemoprevention of cancer or inflammatory diseases.
Assuntos
Isoenzimas/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , NF-kappa B/efeitos dos fármacos , Óxido Nítrico Sintase/efeitos dos fármacos , Prostaglandina-Endoperóxido Sintases/efeitos dos fármacos , Sapogeninas/farmacologia , Triterpenos/farmacologia , Animais , Western Blotting , Células Cultivadas , Ciclo-Oxigenase 2 , Ensaio de Desvio de Mobilidade Eletroforética , Ginsenosídeos/metabolismo , Isoenzimas/biossíntese , Lipopolissacarídeos/farmacologia , Ativação de Macrófagos , Macrófagos/metabolismo , Camundongos , NF-kappa B/metabolismo , Óxido Nítrico Sintase/biossíntese , Óxido Nítrico Sintase Tipo II , Prostaglandina-Endoperóxido Sintases/biossínteseRESUMO
In order to validate the use of the stem bark of Catalpa ovata G. Don. (Bignoniaceae) as an anti-inflammatory drug in the traditional Korean medicine, we have investigated the effects of the methanol extract of this folk medicine on the productions of tumor necrosis factor-alpha (TNF-alpha) and nitric oxide (NO) on RAW 264.7 macrophages activated with the endotoxin lipopolysaccharide. The extract inhibited the productions of TNF-alpha and NO with significant decreases in mRNA levels of TNF-alpha and inducible NO synthase, suggesting that the stem bark of Catalpa ovata may have therapeutic potential in the control of inflammatory disorders.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Bignoniaceae/química , Macrófagos/efeitos dos fármacos , Óxido Nítrico/biossíntese , Fator de Necrose Tumoral alfa/biossíntese , Animais , Anti-Inflamatórios não Esteroides/isolamento & purificação , Western Blotting , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Macrófagos/metabolismo , Camundongos , Casca de Planta/química , Extratos Vegetais/farmacologia , RNA Mensageiro/biossíntese , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Experimental allergic encephalomyelitis (EAE) is an autoimmune CD4+ T cell-mediated disease of the central nervous system (CNS). Nitric oxide (NO) plays an important role in preventing the development of EAE. Molsidomine (Mol) is a drug used for the treatment of coronary artery disease. Its therapeutic effects are the consequences of NO formation. In this study, we investigated the effects of Mol on EAE development in myelin basic protein (MBP)-immunized Lewis rats. All rats immunized with MBP developed typical clinical signs of acute EAE. In the EAE rats receiving Mol, the severity of clinical signs and the infiltration of inflammatory cells in CNS were clearly reduced. Furthermore, Mol administration significantly reduced the production of interferon-gamma, a Th1 inflammatory cytokine, but increased the production of interleukin-10, a Th2 anti-inflammatory cytokine. Our findings suggest that the administration of the exogenous NO donor Mol is of considerable benefit in limiting the development of EAE and other Th1 cell-mediated inflammatory diseases.
Assuntos
Encefalomielite Autoimune Experimental/prevenção & controle , Molsidomina/uso terapêutico , Doadores de Óxido Nítrico/uso terapêutico , Animais , Citocinas/efeitos dos fármacos , Modelos Animais de Doenças , Encefalomielite Autoimune Experimental/induzido quimicamente , Encefalomielite Autoimune Experimental/patologia , Imuno-Histoquímica , Interferon gama/metabolismo , Interleucina-10/metabolismo , Masculino , Molsidomina/farmacologia , Proteína Básica da Mielina , Mielite/patologia , Mielite/fisiopatologia , Doadores de Óxido Nítrico/farmacologia , Ratos , Ratos Endogâmicos LewRESUMO
Recently we have reported that the trichothecene mycotoxin 4-acetyl-12,13-epoxyl-9-trichothecene-3,15-diol (AETD) from the fruiting bodies of Isaria japonica Yasuda is a potent inducer of apoptosis in human promyelocytic HL-60 cells. The present study aims to characterize the molecular events leading to AETD-induced apoptosis in HL-60 cells. The percentage of apoptotic cells (annexin-V-positive cell population) increased dose- and time-dependently after AETD exposure. Apoptosis of HL-60 cells by AETD was associated with the formation of intracellular reactive oxygen species (ROS), the depletion of intracellular glutathione (GSH) and the activation of caspase-3. Pretreating the cells with the antioxidant N-acetyl-L-cystein (NAC) and the caspase-3 inhibitor Z-DEVD-fmk abrogated AETD-induced apoptosis and caspase-3 activation. NAC blocked intracellular ROS formation and GSH depletion, but Z-DEVD-fmk did not. These results indicate that AETD induces apoptosis in HL-60 cells by causing intracellular ROS formation and GSH depletion followed by the downstream event of caspase-3 activation.
Assuntos
Apoptose/efeitos dos fármacos , Ascomicetos/química , Espécies Reativas de Oxigênio/efeitos adversos , Tricotecenos/farmacologia , Caspase 3 , Caspases/farmacologia , Relação Dose-Resposta a Droga , Glutationa/metabolismo , Células HL-60 , HumanosRESUMO
In an effort to elucidate the mechanism of the anti-inflammatory effect of mudanpi, the root cortex of Paeonia suffruticosa Andrews (Ranunculaceae), we determined the effects of the methanolic extract of mudanpi (MEM) on the secretions of interleukin (IL)-8, a major mediator of acute neutrophil-mediated inflammation, and macrophage chemoattractant protein (MCP)-1, a major mediator of chronic macrophage-mediated inflammation, in human monocytic U937 cells stimulated with phorbol myristate acetate (PMA). MEM significantly inhibited PMA-induced secretions of IL-8 and MCP-1 proteins in a dose-dependent manner. The inhibition of these chemokines by MEM was due to its suppression of IL-8 and MCP-1 genes. In addition, 1,2,3,4,6-penta-O-galloyl-beta-D-glucose, one of major constituents isolated from MEM, inhibited PMA-induced secretions of IL-8 and MCP-1 proteins by its suppression of IL-8 and MCP-1 genes. Thus, one possible anti-inflammatory mechanism of mudanpi, an anti-inflammatory Chinese crude drug, may be to inhibit the secretions of inflammatory chemokines.
Assuntos
Anti-Inflamatórios/farmacologia , Quimiocina CCL2/metabolismo , Interleucina-8/metabolismo , Medicina Tradicional do Leste Asiático , Paeonia/química , Extratos Vegetais/farmacologia , Raízes de Plantas/química , Quimiocina CCL2/genética , Ensaio de Imunoadsorção Enzimática , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Interleucina-8/genética , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Acetato de Tetradecanoilforbol/farmacologia , Células U937RESUMO
The amyloid beta-peptide (Abeta)-induced oxidative stress is a well-established pathway of neuronal cell death in Alzheimer's disease (AD). Salidroside, one of the major compounds from the roots of Rhodiola species (Crassulaceae), was investigated in vitro for its cytoprotection against Abeta-induced toxicity on rat neuronal PCl2 cells. Salidroside significantly reduced Abeta-induced cytotoxicity in a dose-dependent manner. Salidroside also reduced Abeta-mediated intracellular accumulation of reactive oxygen species and malondialdehyde (MDA), a product of lipid peroxides, by preventing Abeta-induced decline of antioxidant enzyme activities. These results suggest that salidroside protects neuronal PC12 cells from Abeta-induced cytotoxicity via its antioxidant pathway.
Assuntos
Peptídeos beta-Amiloides/metabolismo , Antioxidantes/farmacologia , Citoproteção , Glucosídeos/farmacologia , Neurônios/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Fenóis/farmacologia , Rhodiola , Animais , Morte Celular , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Glucosídeos/isolamento & purificação , Peroxidação de Lipídeos/efeitos dos fármacos , Malondialdeído/metabolismo , Neurônios/metabolismo , Neurônios/patologia , Células PC12 , Fenóis/isolamento & purificação , Raízes de Plantas , Ratos , Espécies Reativas de Oxigênio/metabolismo , Rhodiola/químicaRESUMO
Catalposide, the major iridoid glycoside isolated from the stem bark of Catalpa ovata G. Don (Bignoniaceae), was found to inhibit the productions of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), and interleukin-6 (IL-6), and the activation of nuclear factor kappaB (NF-kappaB) in RAW 264.7 macrophages activated with lipopolysaccharide (LPS). Catalposide also inhibited the expressions of TNF-alpha, IL-1beta, and IL-6 genes and the nuclear translocation of p65 subunit of NF-kappaB in LPS-activated RAW 264.7 cells. Flow cytometric analysis revealed that catalposide suppressed the binding of FITC-conjugated LPS to CD14 on the surface of cells, probably resulting in the inhibitory effects on TNF-alpha, IL-1beta, and IL-6 productions and NF-kappaB activation. These findings suggest that catalposide could be an attractive candidate for adjunctive therapy in gram-negative bacterial infections.
Assuntos
Bignoniaceae , Glucosídeos/farmacologia , Interleucina-1/antagonistas & inibidores , Interleucina-6/antagonistas & inibidores , Macrófagos/efeitos dos fármacos , NF-kappa B/antagonistas & inibidores , Animais , Bignoniaceae/química , Linhagem Celular , Glucosídeos/química , Glucosídeos/isolamento & purificação , Glicosídeos/química , Glicosídeos/isolamento & purificação , Glicosídeos/farmacologia , Interleucina-1/biossíntese , Interleucina-6/biossíntese , Lipopolissacarídeos/farmacologia , Macrófagos/metabolismo , Camundongos , NF-kappa B/metabolismo , Casca de Planta , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologiaRESUMO
The mitogen-activated protein kinase (MAPK) family members have been implicated in cell survival. We have previously demonstrated that cytotoxic lectin-II isolated from Korean mistletoe induces apoptotic cell death in the human monoblastic leukemia cell line, U937, via the activation of the stress-activated protein kinases/c-Jun N-terminal kinase (SAPK/JNK). In the present study, the roles of extracellular signal-regulated kinases (ERK1/2) and p38 MAPK in lectin-II-induced apoptosis have been investigated. Treatment of U937 cells with lectin-II resulted in apoptotic DNA fragmentation, which was preceded by the activation of ERK1/2, p38 MAPK and SAPK/JNK. This lectin-II-induced DNA fragmentation was significantly enhanced when ERK1/2 activation was selectively inhibited by PD098059. 12-O-tetradecanoylphorbol-13-acetate, which stimulates ERK activity in U937 cells, markedly reduced lectin-II-induced DNA fragmentation. Inhibition of p38 MAPK activity with p38-specific inhibitor, SB203580, partially inhibited lectin-II-induced DNA fragmentation. These results suggest that ERK1/2 and p38 MAPK may have opposite effects on cell survival in response to cytotoxic mistletoe lectin-II, which may contribute to the modulation of lectin-II-mediated cytotoxic activity.
Assuntos
Apoptose/efeitos dos fármacos , Erva-de-Passarinho , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Preparações de Plantas , Proteínas de Plantas , Transdução de Sinais/fisiologia , Toxinas Biológicas/farmacologia , Apoptose/fisiologia , Fragmentação do DNA/efeitos dos fármacos , Flavonoides/farmacologia , Humanos , Imidazóis/farmacologia , Proteínas Quinases JNK Ativadas por Mitógeno , Leucemia Monocítica Aguda/metabolismo , Proteína Quinase 1 Ativada por Mitógeno/efeitos dos fármacos , Proteína Quinase 3 Ativada por Mitógeno , Proteínas Quinases Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases Ativadas por Mitógeno/efeitos dos fármacos , Piridinas/farmacologia , Proteínas Inativadoras de Ribossomos Tipo 2 , Acetato de Tetradecanoilforbol/farmacologia , Células U937/metabolismo , Proteínas Quinases p38 Ativadas por MitógenoRESUMO
Inflammation has been known to be associated with excess synthesis of nitric oxide (NO) by inducible NO synthase (iNOS). Retinoids have been reported to have anti-inflammatory activity, but the mechanism by which they can elicit this activity is poorly understood. The effects of retinoids on NO synthesis and iNOS gene expression in murine fibroblast L929 cells were examined. Treatment of the cells with interferon-y resulted in excess NO synthesis and iNOS gene expression. All-trans-retinoic acid significantly inhibited NO synthesis and iNOS gene expression in a dose-dependent manner. Similarly, 9-cis-retinoic acid also inhibited NO synthesis, but retinol did not show any inhibitory effect on NO synthesis. These findings suggest that the modulation of iNOS gene expression is another possible pathway by which retinoids may function as anti-inflammatory agents.
Assuntos
Óxido Nítrico Sintase/genética , Tretinoína/farmacologia , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Linhagem Celular , Expressão Gênica/efeitos dos fármacos , Inflamação/enzimologia , Inflamação/genética , Interferon gama/toxicidade , Camundongos , Óxido Nítrico/biossíntese , Óxido Nítrico Sintase Tipo II , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas RecombinantesRESUMO
The root of Paeonia suffruticosa ANDREWS is an important Chinese crude drug used in many traditional prescriptions. 1,2,3,4,6-penta-O-galloyl-beta-D-glucose (PGG), a major component of this crude drug, was found to exhibit in vitro growth-inhibiting effect on human hepatocellular carcinoma cell line, SK-HEP-1 cells. The growth-inhibitory effect was related to the ability of PGG not only to cause a G(0)/G(1) phase arrest but also to suppress the activation of nuclear factor-kappa B. Neither apoptosis nor necrosis was observed in the cells treated with PGG. These findings suggest that PGG could be a candidate for developing a low-toxic anticancer agent.
Assuntos
Antineoplásicos/farmacologia , Carcinoma Hepatocelular/patologia , Taninos Hidrolisáveis , Taninos/farmacologia , Apoptose/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , DNA/metabolismo , Ensaio de Desvio de Mobilidade Eletroforética , Fase G1/efeitos dos fármacos , Humanos , NF-kappa B/metabolismo , Necrose , Fase de Repouso do Ciclo Celular/efeitos dos fármacos , Células Tumorais CultivadasAssuntos
Inibidores Enzimáticos/farmacologia , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico Sintase/biossíntese , Fenóis/farmacologia , Psoralea/química , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Northern Blotting , Western Blotting , Linhagem Celular , Núcleo Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Eletroforese , Indicadores e Reagentes , Macrófagos/efeitos dos fármacos , Camundongos , NF-kappa B/efeitos dos fármacos , Óxido Nítrico/biossíntese , Óxido Nítrico Sintase/genética , Óxido Nítrico Sintase Tipo II , Preparações de Plantas/farmacologia , Sementes/químicaRESUMO
Apoptosis is now widely accepted as playing a role in tumorigenesis. An effective compound which can kill tumors via apoptotic pathway appears to be a relevant strategy to suppress various human tumors. The ethyl acetate extract from the stem bark of Cudrania tricuspidata (EACT) showed dose- and time-dependent cytotoxic effects on human leukemia HL-60 cells. DNA fragmentation and morphological changes, accompanied by condensed and fragmented nuclei, were observed in the cells cultured for 6 hr with EACT. These results suggest that the cytotoxicity of the crude extract from Cudrania tricuspidata against HL-60 cells is due to apoptosis.
Assuntos
Antineoplásicos Fitogênicos/antagonistas & inibidores , Apoptose/efeitos dos fármacos , Medicamentos de Ervas Chinesas/farmacologia , Células HL-60/efeitos dos fármacos , Acetatos/farmacologia , Análise de Variância , DNA de Neoplasias/efeitos dos fármacos , Relação Dose-Resposta a Droga , Medicamentos de Ervas Chinesas/uso terapêutico , Eletroforese em Gel de Ágar , Humanos , Técnicas In Vitro , Casca de Planta , Fatores de Tempo , Células Tumorais Cultivadas/efeitos dos fármacosRESUMO
Prunioside A (1) has been isolated from an EtOAc-soluble extract of Spiraea prunifolia var. simpliciflora by a combination of chromatographic techniques. The structure was determined primarily by extensive NMR experiments. Compound 1 is a unique terpene glycoside. Its acetylated derivative (1a) inhibited nitric oxide production in murine macrophage-like RAW 264.7 cells in a dose-dependent manner.
Assuntos
Glicosídeos/isolamento & purificação , Plantas Medicinais/química , Terpenos/isolamento & purificação , Animais , Células Cultivadas/efeitos dos fármacos , Células Cultivadas/metabolismo , Cromatografia Líquida de Alta Pressão , Relação Dose-Resposta a Droga , Glicosídeos/química , Glicosídeos/farmacologia , Coreia (Geográfico) , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Espectroscopia de Ressonância Magnética , Camundongos , Estrutura Molecular , Óxido Nítrico/biossíntese , Óxido Nítrico Sintase/antagonistas & inibidores , Óxido Nítrico Sintase/biossíntese , Óxido Nítrico Sintase Tipo II , Raízes de Plantas/química , Terpenos/química , Terpenos/farmacologiaRESUMO
The fruiting bodies of Isaria fungi have been traditionally used in Korea to treat cancer. An apoptosis-inducing compound, 4-acetyl-12,13-epoxyl-9-trichothecene-3,15-diol, was isolated from the methanol extract of fruiting bodies of Isaria japonica Yasuda by bioassay-guided fractionation. The apoptosis of the human leukemia cells (HL-60) by the compound was accessed by propidium iodide-staining flow cytometric analysis, and apoptosis-inducing activity at IC50 concentration (10 nmol/l) was further confirmed by a nuclear morphological change, a ladder pattern of internucleosomal DNA fragmentation, and an activation of caspase-3.
Assuntos
Apoptose/efeitos dos fármacos , Hypocreales/química , Tricotecenos/farmacologia , Corantes , Fragmentação do DNA/efeitos dos fármacos , DNA de Neoplasias/efeitos dos fármacos , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Células HL-60 , Humanos , Indóis , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Neutrófilos/efeitos dos fármacos , Sais de Tetrazólio , Tiazóis , Tricotecenos/isolamento & purificaçãoRESUMO
In the present study we have investigated whether butein could induce apoptosis in human leukaemic HL-60 cells. The treatment of HL-60 cells with butein induced apoptotic cell death as determined by morphological and biochemical changes. Apoptotic DNA fragments in the butein-treated HL-60 cells were increased gradually as determined by flow cytometric analysis. The caspase-3 activity was increased during butein-induced apoptosis. However, caspase-3 inhibitor abrogated the butein-induced DNA fragmentation. Furthermore, the treatment of HL-60 cells with butein decreased the expression of Bcl-2 protein, but increased the expression of Bax protein. These results suggest that butein-induced apoptosis is mediated through the activation of caspase-3 and it is associated with changed expression of Bcl-2 and Bax proteins.
Assuntos
Apoptose/efeitos dos fármacos , Caspases/metabolismo , Chalcona/análogos & derivados , Chalcona/farmacologia , Inibidores de Fosfodiesterase/farmacologia , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Caspase 3 , Sobrevivência Celular/efeitos dos fármacos , Chalconas , Fragmentação do DNA/efeitos dos fármacos , Relação Dose-Resposta a Droga , Eletroforese em Gel de Ágar , Ativação Enzimática , Precursores Enzimáticos , Citometria de Fluxo , Células HL-60 , Células HeLa , Humanos , Células K562 , Ploidias , Células Tumorais Cultivadas , Células U937 , Proteína X Associada a bcl-2RESUMO
The rhizomes of Cyperus rotundus (C. rotundus) have been used in oriental traditional medicines for the treatment of stomach and bowel disorders, and inflammatory diseases. Nitric oxide (NO) and superoxide (O2-) are important mediators in the pathogenesis of inflammatory diseases. This study was undertaken to address whether the metanol (MeOH) extract of rhizomes of C. rotundus could modulate NO and O2- productions by murine macrophage cell line, RAW 264.7 cells. The MeOH extract of rhizomes of C. rotundus showed the inhibition of NO production in a dose-dependent manner by RAW 264.7 cells stimulated with interferon-gamma plus lipopolysaccharide. The inhibition of NO production by the extract was due to the suppression of iNOS protein, as well as iNOS mRNA expression, determined by Western and Northern blotting analyses, respectively. In addition, the MeOH extract suppressed the production of O2- by phorbol ester-stimulated RAW 264.7 cells in dose- and time-dependent manners. Collectively, these results suggest that the MeOH extract of rhizomes of C. rotundus could be developed as anti-inflammatory candidate for the treatment of inflammatory diseases mediated by overproduction of NO and O2-.