RESUMO
Objective: New treatments are needed for cases of asthma that are refractory to traditional therapies. In this study, we examined the effect of oral nintedanib, an intracellular inhibitor of tyrosine kinases, on airway hyper-responsiveness (AHR) and airway smooth muscle cells, using a mouse model of experimental asthma. Methods: Asthma was experimentally induced in mice via subcutaneous injection of ovalbumin (OVA). A group of saline-injected mice served as a control group. The OVA mice were then divided into four treatment groups according to the dose of nintedanib. AHR was examined via exposure to vaporized methacholine. Airway inflammation was assessed via bronchoalveolar lavage fluid (BALF) cell counts and Th2 cytokine concentrations. Results: Baseline levels of AHR and airway inflammation were higher in OVA mice than in the control group. Treatment with nintedanib lowered AHR, BALF cell counts and BALF cytokine levels in a dose-dependent fashion. The effect of nintedanib was comparable to that of dexamethasone. In particular, treatment with nintedanib lowered the expression of transforming growth factor-ß1 and inhibited the expression and phosphorylation of platelet-derived growth factor receptor-ß, vascular endothelial growth factor receptor 1 (VEGFR1), VEGFR2, fibroblast growth factor receptor 2 (FGFR2), FGFR3, and extracellular signal-regulated kinase. Conclusions: Nintedanib lowered AHR and the expression of factors associated with airway inflammation and remodeling in a mouse model of experimental asthma. Our results suggest that nintedanib may be useful in the treatment of asthma.
Assuntos
Antiasmáticos/administração & dosagem , Asma/tratamento farmacológico , Brônquios/efeitos dos fármacos , Indóis/administração & dosagem , Mediadores da Inflamação/metabolismo , Doença Aguda/terapia , Administração por Inalação , Administração Oral , Remodelação das Vias Aéreas/efeitos dos fármacos , Remodelação das Vias Aéreas/imunologia , Resistência das Vias Respiratórias/efeitos dos fármacos , Resistência das Vias Respiratórias/imunologia , Animais , Asma/diagnóstico , Asma/imunologia , Brônquios/imunologia , Brônquios/metabolismo , Líquido da Lavagem Broncoalveolar/citologia , Broncoconstritores/administração & dosagem , Dexametasona/administração & dosagem , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Feminino , Glucocorticoides/administração & dosagem , Humanos , Mediadores da Inflamação/análise , Cloreto de Metacolina/administração & dosagem , Camundongos , Ovalbumina/administração & dosagem , Ovalbumina/imunologiaRESUMO
In the present study, we demonstrated the anti-catabolic effects of formononetin, a phytoestrogen derived from herbal plants, against interleukin-1ß (IL-1ß)-induced severe catabolic effects in primary rat chondrocytes and articular cartilage. Formononetin did not affect the viability of primary rat chondrocytes in both short- (24 h) and long-term (21 days) treatment periods. Furthermore, formononetin effectively antagonized the IL-1ß-induced catabolic effects including the decrease in proteoglycan content, suppression of pericellular matrix formation, and loss of proteoglycan through the decreased expression of cartilage-degrading enzymes like matrix metalloproteinase (MMP)-13, MMP-1, and MMP-3 in primary rat chondrocytes. Moreover, catabolic oxidative stress mediators like nitric oxide, inducible nitric oxide synthase, cyclooxygenase-2, and prostaglandin E2 were significantly downregulated by formononetin in primary rat chondrocytes treated with IL-1ß. Sequentially, the upregulation of pro-inflammatory cytokines (like IL-1α, IL-1ß, IL-6, and tumor necrosis factor α), chemokines (like fractalkine, monocyte chemoattractant protein-1, and macrophage inflammatory protein-3α), and vascular endothelial growth factor were significantly downregulated by formononetin in primary rat chondrocytes treated with IL-1ß. These data suggest that formononetin may suppress IL-1ß-induced severe catabolic effects and osteoarthritic condition. Furthermore, formononetin may be a promising candidate for the treatment and prevention of osteoarthritis.
Assuntos
Condrócitos/patologia , Antagonismo de Drogas , Inflamação/tratamento farmacológico , Interleucina-1beta/farmacologia , Isoflavonas/farmacologia , Metabolismo/efeitos dos fármacos , Animais , Cartilagem Articular/efeitos dos fármacos , Células Cultivadas , Interleucina-1beta/antagonistas & inibidores , Isoflavonas/antagonistas & inibidores , Osteoartrite/prevenção & controle , Fitoestrógenos/farmacologia , RatosRESUMO
A total of seven phenolics and 44 metabolites was profiled in white flowers of Magnolia denudata and violet flowers of Magnolia liliiflora using high-performance liquid chromatography (HPLC), electrospray ionization-mass spectrometry (ESI-MS), and gas chromatography time-of-flight mass spectrometry (GC-TOFMS). Seven phenylpropanoid compounds were identified in white flowers by liquid chromatography mass spectrometry (LC-MS). An HPLC analysis showed that phenylpropanoid accumulation in violet flowers was 1.48 times higher than that in white flowers. Furthermore, superoxide dismutase (SOD)-like activity and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity were determined to investigate the antioxidant properties of secondary metabolites in different flowers. Violet flowers showed higher SOD-like and DPPH activity than white flowers. In addition, anti-inflammatory activity measured using a nitric oxide assay was higher in violet flowers than in white flowers. Our results provide valuable information on the relationship between primary and secondary metabolites, and synergistic antioxidant and anti-inflammatory properties derived from phenolic compounds in different colored flowers.
Assuntos
Flores/química , Magnolia/química , Antioxidantes/química , Compostos de Bifenilo/química , Cromatografia Líquida de Alta Pressão , Picratos/química , Extratos Vegetais/química , Espectrometria de Massas por Ionização por Electrospray , Superóxido Dismutase/metabolismoRESUMO
Charantin, a natural cucurbitane type triterpenoid, has been reported to have beneficial pharmacological functions such as anticancer, antidiabetic, and antibacterial activities. However, accumulation of charantin in bitter melon has been little studied. Here, we performed a transcriptome analysis to identify genes involved in the triterpenoid biosynthesis pathway in bitter melon seedlings. A total of 88,703 transcripts with an average length of 898 bp were identified in bitter melon seedlings. On the basis of a functional annotation, we identified 15 candidate genes encoding enzymes related to triterpenoid biosynthesis and analyzed their expression in different organs of mature plants. Most genes were highly expressed in flowers and/or fruit from the ripening stages. An HPLC analysis confirmed that the accumulation of charantin was highest in fruits from the ripening stage, followed by male flowers. The accumulation patterns of charantin coincide with the expression pattern of McSE and McCAS1, indicating that these genes play important roles in charantin biosynthesis in bitter melon. We also investigated optimum light conditions for enhancing charantin biosynthesis in bitter melon and found that red light was the most effective wavelength.
Assuntos
Momordica charantia/genética , Extratos Vegetais/metabolismo , Proteínas de Plantas/genética , Triterpenos/metabolismo , Frutas/genética , Frutas/metabolismo , Momordica charantia/química , Momordica charantia/metabolismo , Proteínas de Plantas/metabolismo , Plântula/genética , Plântula/metabolismoRESUMO
Carbohydrate sources play important roles in energy and growth of plants. Therefore, in this study, we investigated the optimal carbohydrate source in hairy root cultures (HRCs) of Scutellaria baicalensis infected with Agrobacterium rhizogenes strain R1000. The hairy roots were cultured in half-strength B5 liquid medium supplemented with seven different carbohydrates sources (sucrose, fructose, glucose, galactose, sorbitol, mannitol and maltose), each at a concentration of 100 mM, in order to identify the best carbon sources for the production of major flavones, such as wogonin, baicalin and baicalein. Sucrose, galactose and fructose markedly influenced the production of major flavones and were therefore chosen for subsequent experiments. HRC growth and flavone accumulation were examined following culture with 30, 100 and 150 mM sucrose, galactose and fructose, respectively. From these data, 150 mM sucrose was found to be the optimal carbon source for the enhancement of baicalein production and growth of S. baicalensis HRCs. Fructose caused the greatest increase in baicalin accumulation. Additionally, galactose was the optimal carbon source for wogonin production. These results provide important insights into the optimal growth conditions, particularly the appropriate carbohydrate source, for S. baicalensis.
Assuntos
Carboidratos/química , Flavonoides/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Scutellaria baicalensis/metabolismo , Metabolismo dos Carboidratos , Células Cultivadas , Meios de Cultura/química , Meios de Cultura/metabolismo , Raízes de Plantas/química , Raízes de Plantas/metabolismo , Scutellaria baicalensis/química , Scutellaria baicalensis/crescimento & desenvolvimentoRESUMO
Esculetin is known to inhibit tumor growth, but its effect in angiogenesis has not been studied. Here, we report the efficacy of esculetin on VEGF-induced angiogenesis. Esculetin treatment inhibited VEGF-induced proliferation and DNA synthesis of HUVECs with no cell toxicity. G1-phase cell-cycle arrest was associated with a decreased expression of cyclins and CDKs via the binding of p27KIP1. Esculetin down-regulated the MMP-2 expression in VEGF-stimulated HUVECs, which suppressed colony tube formation and migration. Esculetin reduced the phosphorylation of VEGFR-2 and the downstream signaling of VEGFR-2, including ERK1/2 and eNOS/Akt pathways. Esculetin suppressed microvessel outgrowth from an aortic ring ex vivo model treated with VEGF, and blocked the VEGF-induced formation of new blood vessels and hemoglobin content in an in vivo Matrigel plug model. Collectively, VEGF-stimulated responses in angiogenesis were inhibited in vitro and in vivo, providing a theoretical basis for effective use against anti-angiogenic therapies.
Assuntos
Antineoplásicos Fitogênicos , Antioxidantes , Neovascularização Patológica/induzido quimicamente , Neovascularização Patológica/tratamento farmacológico , Fitoterapia , Umbeliferonas/farmacologia , Umbeliferonas/uso terapêutico , Fator A de Crescimento do Endotélio Vascular/efeitos adversos , Artemisia , Hemoglobinas/metabolismo , Células Endoteliais da Veia Umbilical Humana , Humanos , Metaloproteinase 2 da Matriz/metabolismo , Fosforilação/efeitos dos fármacos , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismoRESUMO
Radish sprouts and young seedlings are considered important dietary vegetables in Asian countries. In this study, we investigated the levels of glucosinolate and anthocyanin accumulation in radish seedlings in response to light and methyl jasmonate (MeJA) treatments. MeJA facilitated the accumulation of glucosinolate and anthocyanins under light conditions. The glucosinolate and anthocyanin contents in the radish seedlings that were exposed to light after MeJA treatment were higher than those of the seedlings that were grown in the dark without MeJA. At a concentration of 100 µM, MeJA led to the greatest accumulation of the most glucosinolates under both light and dark conditions. Under light conditions, the levels of glucoraphenin, glucoerucin, and glucotropaeolin accumulation were 1.53-, 1.60-, and 1.30-fold higher, respectively, than those of the control. Remarkable accumulations of glucobrassicin were observed under light conditions (4.4-, 6.7-, and 7.8-fold higher than that of the control following the application of 100, 300, and 500 µM MeJA, respectively). The level of cyanidin in the 300 µM MeJA-treated seedlings was double of that in the control without MeJA treatment. The highest level of pelargonidin was observed after treatment with 500 µM MeJA under light conditions; this level was 1.73 times higher than that in the control. A similar trend of anthocyaninaccumulation was observed in the radish seedlings following MeJA treatment under dark conditions, but the levels of anthocyanins were considerably lower in the seedlings that were grown in the dark. Our findings suggest that light and low concentrations of MeJA enhance the accumulations of glucosinolates and anthocyanins during the development of radish seedlings.
Assuntos
Acetatos/metabolismo , Antocianinas/biossíntese , Ciclopentanos/metabolismo , Glucosinolatos/biossíntese , Oxilipinas/metabolismo , Raphanus/metabolismo , Raphanus/efeitos da radiação , Luz , Plântula/metabolismo , Plântula/efeitos da radiaçãoRESUMO
We investigated Licochalcone-A (Lico-A)-induced apoptosis and the pathway underlying its activity in a pharyngeal squamous carcinoma FaDu cell line. Lico-A purified from root of Glycyrrhiza inflata had cytotoxic effects, significantly increasing cell death in FaDu cells. Using a cell viability assay, we determined that the IC50 value of Lico-A in FaDu cells was approximately 100 µM. Chromatin condensation was observed in FaDu cells treated with Lico-A for 24 h. Consistent with this finding, the number of apoptotic cells increased in a time-dependent manner when FaDu cells were treated with Lico-A. TRAIL was significantly up-regulated in Lico-A-treated FaDu cells in a dose-dependent manner. Apoptotic factors such as caspases and PARP were subsequently activated in a caspase-dependent manner. In addition, levels of pro-apoptotic factors increased significantly in response to Lico-A treatment, while levels of anti-apoptotic factors decreased. Lico-A-induced TRAIL expression was mediated in part by a MAPK signaling pathway involving ERK1/2 and p38. In xenograft mouse model, Lico-A treatment effectively suppressed the growth of FaDu cell xenografts by activating caspase-3, without affecting the body weight of mice. Taken together, these data suggest that Lico-A has potential chemopreventive effects and should therefore be developed as a chemotherapeutic agent for pharyngeal squamous carcinoma.
Assuntos
Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Carcinoma de Células Escamosas/genética , Chalconas/farmacologia , Neoplasias de Cabeça e Pescoço/genética , Ligante Indutor de Apoptose Relacionado a TNF/metabolismo , Animais , Caspase 3/genética , Caspase 3/metabolismo , Caspase 7/genética , Caspase 7/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Quimioprevenção , Glycyrrhiza/química , Humanos , Concentração Inibidora 50 , Queratinócitos/efeitos dos fármacos , Queratinócitos/metabolismo , Masculino , Camundongos , Camundongos Nus , Proteína Quinase 3 Ativada por Mitógeno/genética , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Fosforilação , Extratos Vegetais/farmacologia , Raízes de Plantas/química , Transdução de Sinais , Carcinoma de Células Escamosas de Cabeça e Pescoço , Ligante Indutor de Apoptose Relacionado a TNF/genética , Ensaios Antitumorais Modelo de Xenoenxerto , Proteínas Quinases p38 Ativadas por Mitógeno/genética , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
In the present study, we examined the anticancer properties of berberine in KB oral cancer cells with a specific focus on its cellular mechanism. Berberine did not affect the cell viability of the primary human normal oral keratinocytes that were used as a control. However, the viability of KB cells was found to decrease significantly in the presence of berberine in a dose-dependent manner. Furthermore, in KB cells, berberine induced the fragmentation of genomic DNA, changes in cell morphology, and nuclear condensation. In addition, caspase-3 and -7 activation, and an increase in apoptosis were observed. Berberine was also found to upregulate significantly the expression of the death receptor ligand, FasL. In turn, this upregulation triggered the activation of pro-apoptotic factors such as caspase-8, -9 and -3 and poly(ADP-ribose) polymerase (PARP). Furthermore, pro-apoptotic factors such as Bax, Bad and Apaf-1 were also significantly upregulated by berberine. Anti-apoptotic factors such as Bcl-2 and Bcl-xL were downregulated. Z-VAD-FMK, a cell-permeable pan-caspase inhibitor, suppressed the activation of caspase-3 and PARP. These results clearly indicate that berberine-induced cell death of KB oral cancer cells was mediated by both extrinsic death receptor-dependent and intrinsic mitochondrial-dependent apoptotic signaling pathways. In addition, berberine-induced upregulation of FasL was shown to be mediated by the p38 MAPK signaling pathway. We also found that berberine-induced migration suppression was mediated by downregulation of MMP-2 and MMP-9 through phosphorylation of p38 MAPK. In summary, berberine has the potential to be used as a chemotherapeutic agent, with limited side-effects, for the management of oral cancer.
Assuntos
Apoptose/efeitos dos fármacos , Berberina/uso terapêutico , Proteína Ligante Fas/metabolismo , Neoplasias Bucais/tratamento farmacológico , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Fator Apoptótico 1 Ativador de Proteases/metabolismo , Caspase 3/metabolismo , Caspase 7/metabolismo , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Citometria de Fluxo , Humanos , Immunoblotting , Neoplasias Bucais/enzimologia , Neoplasias Bucais/patologia , Proteína X Associada a bcl-2/metabolismo , Proteína de Morte Celular Associada a bcl/metabolismoRESUMO
CONTEXT: Saussurea lappa Dence (Compositae) is used as a traditional herbal medicine to treat abdominal pain and tenesmus in East Asia. Current studies have shown that S. lappa has anticancer activity in divergent of cancer cells. However, the effects of S. lappa on oral cancer and its mechanisms of action have yet to be elucidated. OBJECTIVE: To explore its potential chemotherapeutic effects and mechanism of cell growth inhibition on human oral cancer cells. MATERIALS AND METHODS: The dried roots of S. lappa were used in this study. Cell viability of KB cells was evaluated by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide assay after treatment with 30 µg/ml of methanol extract from the dried roots of S. lappa. To understand whether its effect on cell death is related with apoptosis pathway, we performed DNA fragmentation assay, western blot, caspase activity assay and fluorescence-activated cell sorting (FACS) analysis. RESULTS: Treatment of S. lappa extract onto KB cells reduced cell viability significantly with an IC50 value of 30 µg/ml. The formation of a DNA ladder was observed starting at the 24 h treatment. In western blotting analysis, the S. lappa extract induced the proteolytic processing of caspase-3, -9 and poly (ADP-ribose) polymerase, a significant increase of Bax and marked reduction of Bcl-2. We also confirmed the activation of caspase-3/-7 in living KB cells by fluorescence microscopy. CONCLUSION: These results suggested that S. lappa extract inhibited cell proliferation through the apoptosis pathway in KB human oral cancer cells.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Neoplasias Bucais/patologia , Extratos Vegetais/farmacologia , Saussurea , Antineoplásicos Fitogênicos/isolamento & purificação , Caspase 3/metabolismo , Caspase 7/metabolismo , Caspase 9/metabolismo , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Fragmentação do DNA , Relação Dose-Resposta a Droga , Humanos , Células KB , Metanol/química , Neoplasias Bucais/metabolismo , Fitoterapia , Extratos Vegetais/isolamento & purificação , Raízes de Plantas , Plantas Medicinais , Poli(ADP-Ribose) Polimerases/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Saussurea/química , Transdução de Sinais/efeitos dos fármacos , Solventes/química , Fatores de Tempo , Proteína X Associada a bcl-2/metabolismoRESUMO
Common buckwheat (Fagopyrum esculentum Moench) is rich in phenolic compounds and may be useful for the treatment of metabolic syndrome in humans. To improve the production of rutin in buckwheat, we overexpressed the flavonol-specific transcription factor, AtMYB12 using Agrobacterium rhizogenes into hairy root culture systems. This induced the expression of flavonoid biosynthetic genes encoding phenylalanine ammonia lyase, cinnamate 4-hydroxylase, 4-coumarate:CoA ligase, chalcone synthase, chalcone isomerase, flavone 3-hydroxylase, flavonoid 3'-hydroxylase, and flavonol synthase. This led to the accumulation of rutin in buckwheat hairy roots up to 0.9 mg/g dry wt. PAP1 expression, however, did not correlate with the production of rutin.
Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Fagopyrum/metabolismo , Engenharia Metabólica/métodos , Raízes de Plantas/metabolismo , Rutina/metabolismo , Fatores de Transcrição/genética , Agrobacterium/genética , Técnicas de Cultura de Células/métodos , Fagopyrum/genética , Expressão Gênica , Proteínas Associadas a Pancreatite , Raízes de Plantas/genética , Proteínas Recombinantes/genéticaRESUMO
Phytoene synthase (PSY) and phytoene desaturase (PDS), which catalyze the first and second steps of the carotenoid biosynthetic pathway, respectively, are key enzymes for the accumulation of carotenoids in many plants. We isolated 2 partial cDNAs encoding PSY (AsPSY-1 and AsPSY-2) and a partial cDNA encoding PDS (AsPDS) from Allium sativum. They shared high sequence identity and conserved motifs with other orthologous genes. Quantitative real-time PCR analysis was used to determine the expression levels of AsPSY1, AsPSY2, and AsPDS in the bulbils, scapes, leaves, stems, bulbs, and roots of garlic. High-performance liquid chromatography demonstrated that carotenoids were not biosynthesized in the underground organs (roots and bulbs), but were very abundant in the photosynthetic organs (leaves) of A. sativum. A significantly higher amount of ß-carotene (73.44 µg·g(-1)) was detected in the leaves of A. sativum than in the other organs.
Assuntos
Alquil e Aril Transferases/genética , Carotenoides/biossíntese , DNA de Plantas/análise , Alho/enzimologia , Oxirredutases/genética , Alquil e Aril Transferases/química , Sequência de Aminoácidos , Carotenoides/análise , Clonagem Molecular , DNA Complementar/análise , Alho/química , Expressão Gênica , Geranil-Geranildifosfato Geranil-Geraniltransferase , Dados de Sequência Molecular , Oxirredutases/química , Folhas de Planta/enzimologia , Raízes de Plantas/enzimologia , Caules de Planta/enzimologia , Reação em Cadeia da PolimeraseRESUMO
In general, an antimicrobial test for screening anti-caries natural extracts was performed by measuring the minimum bactericidal concentration (MBC) against the type strains of mutans streptococci. However, it is unclear if the antimicrobial efficiency of natural extracts on the type strains of mutans streptococci is the same on the clinical strains. In this study, we introduced a bacterial model system for the screening of anti-caries and determining the optimal concentration of them to develop oral hygiene products for Korean populations.
Assuntos
Antibacterianos/farmacologia , Produtos Biológicos/farmacologia , Cárie Dentária/microbiologia , Avaliação Pré-Clínica de Medicamentos/métodos , Streptococcus mutans/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacosRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Cordyceps militaris, one of traditional herbal ingredient in oriental medicine, has been known to promote anticancer and immunomodulatory activities in vitro and in vivo. However, the biological mechanism of anticancer activity has been unknown. OBJECTIVE: To investigate the effect of Cordyceps militaris extract on expression of interferon gamma (IFN-gamma) through interlukin-18 (IL-18) induction and its biological mechanism in vitro and in vivo. MATERIALS AND METHODS: Mice were administrated orally with solution extracted from Cordyceps militaris. The transcription level of IL-18 and IFN-gamma production were measured by reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry. RAW 264.7 cells were transiently transfected with pCATp1 and pCATp2 for IL-18 promoter functional analysis. RESULTS: Cordyceps militaris extracts treatment significantly induced level of IL-18 transcription in mouse brain and liver and enhanced IL-18 transcription level and activated the IFN-gamma production in RAW 264.7 cells. Furthermore, Cordyceps militaris extract led to increase the activity of pCATp1 construct containing the 5' franking region of IL-18 promoter in RAW 264.7 cells. CONCLUSION: Cordyceps militaris extract induced IL-18 mRNA level via enhancing of P1 promoter region result in activation of IFN-gamma production, indicating its potential as an immune activator or anticancer drug.
Assuntos
Cordyceps , Regulação Fúngica da Expressão Gênica/efeitos dos fármacos , Interferon gama/biossíntese , Interleucina-18/biossíntese , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/imunologia , Cordyceps/genética , Cordyceps/metabolismo , Imunoquímica , Interferon gama/genética , Interleucina-18/genética , Fígado/efeitos dos fármacos , Fígado/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Regiões Promotoras Genéticas/efeitos dos fármacos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , TransfecçãoRESUMO
Carnosic acid (CA) is one of the bioactive polyphenols present in extracts of the herb rosemary (Rosmarinus officinalis). In this study, we examined possible protective effects of CA on neurotoxicity induced by dieldrin, an organochlorine pesticide implicated in sporadic Parkinson's disease, in cultured dopaminergic cells (SN4741). CA (5-10 muM) pretreatment showed potent protective effects in a concentration-related manner and prevented dieldrin (10 muM)-induced caspase-3 activation, Jun N-terminal kinase phosphorylation, and caspase-12 activation. Furthermore, dieldrin-induced downregulation of brain-derived neurotrophic factor production was significantly attenuated by CA. These results suggest that CA may safeguard dopaminergic neuronal cells from environmental neurotoxins by enhancing brain-derived neurotrophic factor and repressing apoptotic molecules.
Assuntos
Abietanos/farmacologia , Apoptose/efeitos dos fármacos , Dieldrin/toxicidade , Dopamina/metabolismo , Neurônios/efeitos dos fármacos , Extratos Vegetais/farmacologia , Animais , Antioxidantes/farmacologia , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Caspase 12/metabolismo , Caspase 3/metabolismo , Linhagem Celular , Relação Dose-Resposta a Droga , Ensaio de Imunoadsorção Enzimática , Immunoblotting , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Neurônios/citologia , Neurônios/metabolismo , Rosmarinus/químicaRESUMO
Buckwheat (Fagopyrum esculentum Moench.) is a potentially important source of rutin, a natural flavonoid with antihyperglycemic, antihypertensive, and antioxidative properties. To examine in vitro production of rutin, we established a hairy root culture of buckwheat by infecting leaf explants with Agrobacterium rhizogenes R1000, and tested the growth conditions and rutin production rates of these cultures. Ten hairy root clones were established; their growth and rutin production rates ranged from 233 to 312 (mg dry wt per 30 mL flask, and 0.8 to 1.2 (mg/g dry wt), respectively. Clone H8, which had high growth and rutin production rates (312 mg dry wt per 30 mL flask and 1.2 mg/g dry wt, respectively), was selected for further experiments. H8 showed maximal growth and rutin content at 30 days in culture in MS medium. Of four tested culture media, half-strength MS medium was found to induce the highest levels of growth (378 mg dry wt per 30 mL flask) and rutin production (1.4 mg/g dry wt) by clone H8. In contrast, supplementation with auxins (0.1-1 mg/l IAA, IBA and NAA) increased the growth rate, but had no significant effect on rutin production by H8. Collectively, these findings indicate that hairy root cultures of buckwheat culture could be a valuable alternative approach for rutin production.
Assuntos
Fagopyrum/crescimento & desenvolvimento , Raízes de Plantas/crescimento & desenvolvimento , Plantas Geneticamente Modificadas/metabolismo , Rutina/biossíntese , Rutina/isolamento & purificação , Técnicas de Cultura de Tecidos , Cromatografia Líquida de Alta Pressão/métodos , Meios de Cultura/química , Extratos Vegetais/química , Raízes de Plantas/química , Plantas Medicinais/química , Rhizobium/enzimologia , Rutina/químicaRESUMO
The aim of this investigation was to determine the content of total phenolics, antioxidant activity and cytotoxicity of methanol extracts from the aerial parts of 11 Korean medicinal salad plants. The highest total phenolic content of the methanol extracts was found in Aster scaber (17.1 mg 100 g(-1)), followed by Ixeris dentate (16.4 mg 100 g(-1)), Aster yomena (12.0 mg 100 g(-1)) and Sedum sarmentosum (9.1 mg 100 g(-1)) of FW. Methanol extracts of Ixeris dentate and Aster scaber at 50 microg mL(-1) exhibited the highest DPPH radical scavenging activity by 86.4 and 83.3%, respectively. It was registered a dose-dependent increase of DPPH free radical scavenging activity. Total phenolic content of the studied plant extracts was correlated with the DPPH radical scavenging activity. It was found by means of MTT assay, that cytotoxicity of the methanol extracts was the highest against HCT-116. Methanol extracts from Petasites japonicus (IC(50)<25.0 microg mL(-1)) showed the highest activity against HCT-116, following by Angelica gigas (34.75 microg mL(-1)), Erythronium japonicum (44.06 microg mL(-1)), and Aster scaber (54.87 microg mL(-1)). In conclusion, the studied salad plants have high total phenolics content and high antioxidant activity. These plants dose-dependently increased DPPH free radical scavenging activity. The total phenolics level was highly correlated with the free radical scavenging activity. Most of the studied salad plants have potent cytotoxicity activity. The results of this investigation suggest that the extracts of studied salad plants could be an addition to basic medicine for some diseases.