RESUMO
Nanomedicine has become a promising approach to improve cancer chemotherapy. It remains a major challenge how to enhance anti-drug efficacy and reduce side effects of anti-cancer drugs. Herein, we report a self-assembled nanoplatform (FDEP NPs) by integration of doxorubicin (DOX) and epigallocatechin-3-O-gallate (EGCG) with the help of coordination between Fe3+ ions and polyphenols. The EGCG from FDEP NPs could inhibit the expression of carbonyl reductase 1 (CBR1) protein and thereby inhibit the doxorubicinol (DOXOL) generation from DOX both in vitro and in vivo, thus the efficacy of DOX to cancerous cells is improved significantly. More importantly, the FDEP NPs could reduce cardiac toxicity and the DOX mediated toxicity to blood cells due to the repression of DOXOL production. Moreover, the blood half-life of FDEP NPs is longer than 23â¯h as determined by positron emission tomography (PET) imaging of biodistribution of radiolabelled NPs and HPLC measurement of plasma level of DOX, ensuring high tumor accumulation of FDEP NPs by enhanced permeability and retention (EPR) effect. The FDEP NPs also exhibited much improved antitumor effect over free drugs. Our work sheds new light on the engineering of nanomaterials for combination chemotherapy and may find unique clinical applications in the near future.
Assuntos
Carbonil Redutase (NADPH)/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Nanoestruturas/química , Polifenóis/farmacologia , Chá/química , Animais , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Carbonil Redutase (NADPH)/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Doxorrubicina/análogos & derivados , Doxorrubicina/farmacologia , Liberação Controlada de Fármacos , Endocitose/efeitos dos fármacos , Inibidores Enzimáticos/farmacocinética , Células HEK293 , Humanos , Camundongos Endogâmicos BALB C , Imagem Óptica , Tomografia por Emissão de Pósitrons , Distribuição Tecidual/efeitos dos fármacosRESUMO
BACKGROUND: Compound Fengshiding capsule (CFC), is a Chinese formulation from herbal origin including Alangium platanifolium, Angelicae dahurica, Cynanchum paniculatum and Glycyrrhiza uralensis. CFC is widely used as clinical therapy against rheumatoid arthritis. However, its exact mechanism of action has not been explored yet. METHODS: In order to explore the synergistic mechanism of CFC, we designed a study adopting network pharmacology scheme to screen the action targets in relation to the CFC components. The study analyses target facts of salicin, paeonol, liquiritin and imperatorin from PubMed database, and explores the potential pharmacological targets of rheumatoid arthritis, cervical neuralgia and sciatica related diseases for their interaction. RESULTS: The results of boosted metabolic pathway showed that the chemical components of CFC interrupted many immune-related pathways, thus participating in immunity regulation of the body and playing a role in the treatment of rheumatism. Collectively, CFC has apoptotic, oxidative stress modulatory and anti-inflammatory effects that accumulatively serve for its clinical application against rheumatoid arthritis. CONCLUSION: Conclusively, our findings from present study reconnoiters and compacts systematic theoretical approach by utilizing the network pharmacology mechanism of four effective components for the treatment of rheumatism indicating sufficient potential drug targets associated with CFC against rheumatism. These interesting findings entreaties for further in vitro and in vivo studies on the mechanism of compound active ingredient against rheumatism.
Assuntos
Artrite Reumatoide/tratamento farmacológico , Medicamentos de Ervas Chinesas/farmacologia , Medicamentos de Ervas Chinesas/uso terapêutico , Artrite Reumatoide/etiologia , Artrite Reumatoide/metabolismo , Biomarcadores , Biologia Computacional/métodos , Bases de Dados Genéticas , Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/química , Ontologia Genética , Humanos , Medicina Tradicional Chinesa , Estrutura Molecular , Mapeamento de Interação de Proteínas , Transdução de Sinais/efeitos dos fármacosRESUMO
Among the various treatments, induction of synoviocyte apoptosis by natural products during a rheumatoid arthritis (RA) pathological condition can be considered to have vast potential. However, it is unclear that liquiritin, a kind of natural flavonoid extracted from the roots of Glycyrrhiza uralensis, induced the apoptosis of the synovial membrane and its molecular mechanism. In this study, interleukin-1ß (IL-1ß)-RA-FLS cells were incubated with different concentrations of liquiritin. An MTT assay, Hoechst 33342 staining, JC-1 staining, and Western blot were used to check the viability, cell apoptosis, mitochondrial membrane potential changes, and the expression of related proteins, respectively. In vivo, a TUNEL assay and HE staining of tissue were used for histopathological evaluation. Our results showed that liquiritin significantly inhibited the proliferation of IL-1ß-induced-RA-FLS, promoted nuclear DNA fragmentation, and changed the mitochondrial membrane potential to accelerate cell apoptosis. Liquiritin downregulated the ratio of Bcl-2/Bax and inhibited the VEGF expression and phosphorylation of JNK and P38. Moreover, liquiritin improved the clinical score of rheumatism, inflammatory infiltration, and angiogenesis and induced apoptosis of the synovial tissue in vivo. Hence, liquiritin ameliorates RA by reducing inflammation, blocking MAPK signaling, and restraining angiogenesis.
Assuntos
Artrite Reumatoide/tratamento farmacológico , Artrite Reumatoide/imunologia , Medicamentos de Ervas Chinesas/administração & dosagem , Flavanonas/administração & dosagem , Glucosídeos/administração & dosagem , Glycyrrhiza uralensis/química , Neovascularização Patológica/tratamento farmacológico , Animais , Apoptose/efeitos dos fármacos , Artrite Reumatoide/genética , Artrite Reumatoide/fisiopatologia , Proliferação de Células/efeitos dos fármacos , Humanos , Interleucina-1beta/genética , Interleucina-1beta/imunologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Masculino , Neovascularização Patológica/genética , Neovascularização Patológica/imunologia , Neovascularização Patológica/fisiopatologia , Fosforilação/efeitos dos fármacos , Ratos , Ratos Wistar , Membrana Sinovial/efeitos dos fármacos , Membrana Sinovial/imunologia , Proteínas Quinases p38 Ativadas por Mitógeno/genética , Proteínas Quinases p38 Ativadas por Mitógeno/imunologiaRESUMO
Rheumatoid arthritis (RA) is a chronic inflammatory disorder linked to oxidative stress of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLSs). The effects and potential mechanism of salicin on inflammation and oxidative stress of RA-FLSs were examined by MTT, ELISA, and Western blot methods. Salicin significantly reduced cell viability (82.03 ± 7.06, P < 0.01), cytokines (47.70 ± 1.48 ng/L for TNF-α, 30.03 ± 3.49 ng/L for IL-6) ( P < 0.01), and matrix metalloproteinases-1/-3 expression ( P < 0.01) in IL-1ß-induced RA-FLSs and inhibited ROS generation and p65 phosphorylation ( P < 0.01) as compared with IL-1ß-induced treatment. Moreover, salicin promoted Nrf2 nuclear translocation (2.15 ± 0.21) and HO-1 expression (1.12 ± 0.05) and reduced ROS production in IL-1ß-induced RA-FLSs ( P < 0.01). Salicin not only reduced the collagen-induced arthritis by reducing the clinical score ( P < 0.01), inflammatory infiltration, and synovial hyperplasia in vivo but also suppressed the oxidative damage indexes (SOD 155.40 ± 6.53 U/mg tissue, MDA 152.80 ± 5.89 nmol/g tissue, GSH 50.98 ± 3.45 nmol/g tissue, and CAT 0.92 ± 0.10 U/g protein) ( P < 0.01) of ankle joint cells. Conclusively, our findings indicate that salicin ameliorates rheumatoid arthritis, which may be associated with oxidative stress and Nrf2-HO-1-ROS pathways in RA-FLSs.
Assuntos
Alangiaceae/química , Artrite Reumatoide/tratamento farmacológico , Álcoois Benzílicos/administração & dosagem , Glucosídeos/administração & dosagem , Heme Oxigenase-1/metabolismo , Proteínas de Membrana/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Extratos Vegetais/administração & dosagem , Espécies Reativas de Oxigênio/metabolismo , Animais , Artrite Reumatoide/genética , Artrite Reumatoide/metabolismo , Heme Oxigenase-1/genética , Humanos , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Proteínas de Membrana/genética , Fator 2 Relacionado a NF-E2/genética , NF-kappa B/genética , NF-kappa B/metabolismo , Ratos , Ratos Wistar , Transdução de Sinais/efeitos dos fármacos , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Heat stress can induce the cultured microspores into embryogenesis. In this study, whole genome bisulphite sequencing was employed to study global DNA methylation variations after short-term heat shock (STHS) treatments in cultured microspores of Brassica napus cv. Topas. Our results indicated that treatment on cultured Topas microspores at 32 °C for 6 h triggered DNA hypomethylation, particularly in the CG and CHG contexts. And the total number of T32 (Topas 32 °C for 6 h) vs. T0 (Topas 0 h) differentially methylated region-related genes (DRGs) was approximately two-fold higher than that of T18 (Topas 18 °C for 6 h) vs. T0 DRGs, which suggested that 32 °C might be a more intense external stimulus than 18 °C resulting in more changes in the DNA methylation status of cultured microspores. Additionally, 32 °C treatment for 6 h led to increased CHG differential methylations of transposons (DMTs), which were mainly constituted by overlaps between the hypomethylated differentially methylated regions (hypo-DMRs) and transposon elements (TEs). Further analysis demonstrated that the DRGs and their paralogs exhibited differential methylated/demethylated patterns. To summarize, the present study is the first methylome analysis of cultured microspores in response to STHS and may provide valuable information on the roles of DNA methylation in heat response.
Assuntos
Brassica napus/genética , Metilação de DNA , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Pólen/genética , Brassica napus/metabolismo , Mapeamento Cromossômico , Cromossomos de Plantas/química , Cromossomos de Plantas/metabolismo , Elementos de DNA Transponíveis , Ontologia Genética , Loci Gênicos , Resposta ao Choque Térmico/genética , Temperatura Alta , Anotação de Sequência Molecular , Proteínas de Plantas/metabolismo , Pólen/metabolismoRESUMO
Extract of Rabdosia amethystoides (Benth) Hara (ERA), a traditional Chinese medicine has antibacterial, antiviral, anti-tumor, anti-hepatitis and anti-inflammatory properties. However, the hepatoprotective effects and molecular mechanisms of ERA on acute liver injury have not been fully elucidated. This study aims to investigate the anti-inflammatory effect and liver protection of ERA against the acute liver injury induced by Concanavalin A (Con A) and its underlying molecular mechanisms in mice. Mice received ERA (50, 100, 150 mg/kg body weight) by gavage before Con A intravenous administration. We found that ERA pretreatment was able to significantly reduce the elevated serum alanine and aspartate aminotransferase levels and liver necrosis in Con A-induced hepatitis. In addition, ERA treatment significantly decreased the myeloperoxidase, malondialdehyde levels and augmented superoxide dismutase level in the liver tissue, and also suppressed the secretion of proinflammatory cytokines in the serum, compared with Con A group by enzyme linked immunosorbent assay. Furthermore, we observed that ERA pretreatment can significantly decrease the expression level of Toll-like receptor (TLR) 4 mRNA or protein in liver tissues. Further results showed that ERA pretreatment was capable of attenuating the activation of the NF-κB pathway by inhibiting IκBα kinase and p65 phosphorylation in Con A-induced liver injury. Our results demonstrate that ERA pretreatment has hepatoprotective property against Con A-induced liver injury through inhibition of inflammatory mediators in mice. The beneficial effect of ERA may be mediated by the downregulation of TLR4 expression and the inhibition of NF-κB activation.
Assuntos
Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle , Concanavalina A/efeitos adversos , Isodon/química , Fígado/metabolismo , NF-kappa B/metabolismo , Extratos Vegetais/farmacologia , Receptor 4 Toll-Like/metabolismo , Animais , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Doença Hepática Induzida por Substâncias e Drogas/genética , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/genética , Expressão Gênica/efeitos dos fármacos , Masculino , Camundongos Endogâmicos ICR , Fitoterapia , Extratos Vegetais/administração & dosagem , Extratos Vegetais/uso terapêutico , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Receptor 4 Toll-Like/genéticaRESUMO
Association mapping can quickly and efficiently dissect complex agronomic traits. Rapeseed is one of the most economically important polyploid oil crops, although its genome sequence is not yet published. In this study, a recently developed 60K Brassica Infinium(®) SNP array was used to analyse an association panel with 472 accessions. The single-nucleotide polymorphisms (SNPs) of the array were in silico mapped using 'pseudomolecules' representative of the genome of rapeseed to establish their hypothetical order and to perform association mapping of seed weight and seed quality. As a result, two significant associations on A8 and C3 of Brassica napus were detected for erucic acid content, and the peak SNPs were found to be only 233 and 128 kb away from the key genes BnaA.FAE1 and BnaC.FAE1. BnaA.FAE1 was also identified to be significantly associated with the oil content. Orthologues of Arabidopsis thaliana HAG1 were identified close to four clusters of SNPs associated with glucosinolate content on A9, C2, C7 and C9. For seed weight, we detected two association signals on A7 and A9, which were consistent with previous studies of quantitative trait loci mapping. The results indicate that our association mapping approach is suitable for fine mapping of the complex traits in rapeseed.
Assuntos
Brassica napus/genética , Mapeamento Cromossômico , Genes de Plantas , Sementes/genética , Proteínas de Arabidopsis/genética , Brassica napus/química , Simulação por Computador , Ácidos Erúcicos/análise , Estudo de Associação Genômica Ampla/métodos , Glucosinolatos/análise , Histona Acetiltransferases/genética , Desequilíbrio de Ligação , Fenótipo , Óleos de Plantas/análise , Polimorfismo de Nucleotídeo Único , Sementes/anatomia & histologiaRESUMO
l-Arginine (l-Arg) supplementation has been shown to enhance physical exercise capacity and delay onset of fatigue. This work investigated the potential beneficial mechanism(s) of l-Arg supplementation by examining its effect on the cellular oxidative and nitrosative stress pathways in the exercised rats. Forty-eight rats were randomly divided into six groups: sedentary control; sedentary control with l-Arg treatment; endurance training (daily swimming training for 8 wk) control; endurance training with l-Arg treatment; an exhaustive exercise (one time swimming to fatigue) control; and an exhaustive exercise with l-Arg treatment. l-Arg (500 mg/kg body wt) or saline was given to rats by intragastric administration 1 h before the endurance training and the exhaustive swimming test. Expression levels and activities of the l-Arg/nitric oxide (NO) pathway components and parameters of the oxidative stress and antioxidant defense capacity were investigated in l-Arg-treated and control rats. The result show that the l-Arg supplementation completely reversed the exercise-induced activation of NO synthase and superoxide dismutase, increased l-Arg transport capacity, and increased NO and anti-superoxide anion levels. These data demonstrate that l-Arg supplementation effectively reduces the exercise-induced imbalance between oxidative stress and antioxidant defense capacity, and this modulation is likely mediated through the l-Arg/NO pathways. The findings of this study improved our understanding of how l-Arg supplementation prevents elevations of reactive oxygen species and favorably enhances the antioxidant defense capacity during physical exercise.
Assuntos
Antioxidantes/farmacologia , Aorta Abdominal/efeitos dos fármacos , Arginina/farmacologia , Suplementos Nutricionais , Músculo Esquelético/efeitos dos fármacos , Óxido Nítrico/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Esforço Físico , Transdução de Sinais/efeitos dos fármacos , Sistemas de Transporte de Aminoácidos Básicos/metabolismo , Animais , Antioxidantes/metabolismo , Aorta Abdominal/metabolismo , Arginina/metabolismo , Ácido Láctico/sangue , Masculino , Fadiga Muscular/efeitos dos fármacos , Músculo Esquelético/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Condicionamento Físico Animal , Ratos , Ratos Sprague-Dawley , Comportamento Sedentário , Superóxido Dismutase/metabolismo , Natação , Fatores de Tempo , Tirosina/análogos & derivados , Tirosina/metabolismoRESUMO
The present study investigated the neuroprotective effects of aucubin on hydrogen peroxide (H2O2)-induced apoptosis in PC12 cells. Exposure of PC12 cells to 0.25 mm H2O2 induced a leakage of lactate dehydrogenase and decreased cell viability, as shown by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. In a dose over 0.1 mm, aucubin increased PC12 cellular viability and markedly attenuated H2O2-induced apoptotic cell death. Quantitation of apoptosis by flow cytometry indicated that aucubin inhibited H2O2-induced apoptosis in PC12 cells. Nuclear damage was alleviated by aucubin, as shown by Hoechst staining. In addition, the levels of malondialdehyde were reduced and the activity of superoxide dismutase, catalase and glutathione peroxidase was augmented in these cells. These results indicated that aucubin inhibited H2O2-induced apoptosis in PC12 cells through regulation of the endogenous oxidant-antioxidant balance. Our results suggest that aucubin is a potential protective agent for the treatment of oxidative-stress-induced neurodegenerative disease.
Assuntos
Apoptose , Peróxido de Hidrogênio/efeitos adversos , Glucosídeos Iridoides/farmacologia , Animais , Antioxidantes/metabolismo , Catalase/metabolismo , Forma do Núcleo Celular , Sobrevivência Celular , Ativação Enzimática , Citometria de Fluxo , Glutationa Peroxidase/metabolismo , Peróxido de Hidrogênio/metabolismo , L-Lactato Desidrogenase/metabolismo , Malondialdeído/metabolismo , Fármacos Neuroprotetores/farmacologia , Estresse Oxidativo , Células PC12 , Ratos , Coloração e Rotulagem , Superóxido Dismutase/metabolismoRESUMO
Accurately quantitative determination of oil content in oilseed rape plays an important role in varieties breeding for improving oil content in seeds. However, large quantity of oilseeds were needed in order to obtain accuracy and precision results by using standard Soxhlet extraction method, which may be a handicap in analysis of small, rare and precious samples in plant breeding. In the present work, ultrasound-assisted extraction was evaluated as a simpler and more effective alternative to conventional extraction method for the isolation of oil from small quantity of oilseed rape (<20 mg). The oil of oilseed rape samples was extracted by ultrasound-assisted method, and then the fatty acids and total oil content of the seeds were qualitatively and quantitatively determined by gas chromatography (GC). Extraction efficiency of total oil obtained by ultrasound-assisted extraction through an orthogonal experiment (L(9) (3(4))) were investigated to get the best extraction conditions. Statistical analysis showed that the variable with the largest effect was the ultrasound-assisted extraction time which was followed by the ultrasound-assisted extraction power, and the liquid:solid ratio. A liquid:solid ratio of 1:4 (L:g), an ultrasound-assisted extraction time of 60 min and an ultrasound-assisted extraction power of 500 W were found to be optimal for oil extraction from oilseed rape. By comparing with the conventional method, it was found that the ultrasound-assisted extraction of oil from oilseed rape was about five times faster than the traditional extraction method. By the use of ultrasound-assisted extraction combined with GC analysis, the fatty acids and total oil content in small quantity of seeds (<20 mg) were successfully qualitatively determined and the results are in agreement with that obtained by traditional standard method.
Assuntos
Brassica rapa/química , Óleos de Plantas/análise , Análise de Variância , Brassica rapa/efeitos da radiação , Cromatografia Gasosa , Ácidos Graxos/análise , Ácidos Graxos Monoinsaturados , Indicadores e Reagentes , Óleos de Plantas/efeitos da radiação , Óleo de Brassica napus , UltrassomRESUMO
A simple, accurate and rapid spectrophotmeric method was proposed for the determination of chondroitin sulfate. The method was based on the absorbances of AA-CS complex at 625 nm being in proportion to the chondroitin sulfate concentrations. The linear range was 0-30 micrograms.mL-1 (R = 0.999), and the recovery range was 97.4%-103.8%. The quantities of chondroitin sulfate in different sample were determined in this way.