RESUMO
During the screening of the cytotoxicity of rare Korean endemic plants, the extract of Thuja koraiensis Nakai displayed potent cytotoxicity against the adenocarcinomic human alveolar basal epithelial A549 cell line. Through a series of separations via column chromatography, three undescribed abietanes, an undescribed labdane along with a labdane, and a biflavonoid were purified from methylene chloride (CH2Cl2) fraction possessing a potent cytotoxic effect. Extensive 1D and 2D NMR spectroscopic data analyses, in combination with quantum chemical calculations were conducted to establish the planar and absolute configurations of thujakoraienes A-C. The chemical structure of thujakoraiene D was elucidated by spectroscopic data analysis and competing enantioselective acylation. Thujakoraienes A and C along with 7,7â³-di-O-methylamentoflavone, showed cytotoxic effects on A549 cells, with IC50 values of 64.86, 47.97, and 16.14 µM, respectively. Finally, thujakoraiene C and 7,7â³-di-O-methylamentoflavone were identified as potent cytotoxic compounds in A549 cells, followed by an additional cytotoxicity test in the normal human lung fibroblast MRC-5 cell line. This is the first study on the non-volatile chemicals in the extract of T. koraiensis and comparison of chemical profiles of T. orientalis and T. koraiensis.
Assuntos
Antineoplásicos , Diterpenos , Thuja , Humanos , Células A549 , Thuja/química , Estrutura Molecular , Antineoplásicos/farmacologia , Diterpenos/química , Extratos Vegetais/farmacologia , Linhagem Celular TumoralRESUMO
Memory deterioration in Alzheimer's disease (AD) is thought to be underpinned by aberrant amyloid ß (Aß) accumulation, which contributes to synaptic plasticity impairment. Avenanthramide-C (Avn-C), a polyphenol compound found predominantly in oats, has a range of biological properties. Herein, we performed methanolic extraction of the Avns-rich fraction (Fr. 2) from germinated oats using column chromatography, and examined the effects of Avn-C on synaptic correlates of memory in a mouse model of AD. Avn-C was identified in Fr. 2 based on 1H-NMR analysis. Electrophysiological recordings were performed to examine the effects of Avn-C on the hippocampal long-term potentiation (LTP) in a Tg2576 mouse model of AD. Avn-C from germinated oats restored impaired LTP in Tg2576 mouse hippocampal slices. Furthermore, Avn-C-facilitated LTP was associated with changes in the protein levels of phospho-glycogen synthase kinase-3ß (p-GSK3ß-S9) and cleaved caspase 3, which are involved in Aß-induced synaptic impairment. Our findings suggest that the Avn-C extract from germinated oats may be beneficial for AD-related synaptic plasticity impairment and memory decline.
Assuntos
Doença de Alzheimer/tratamento farmacológico , Hipocampo/efeitos dos fármacos , Potenciação de Longa Duração/efeitos dos fármacos , ortoaminobenzoatos/farmacologia , Doença de Alzheimer/metabolismo , Doença de Alzheimer/patologia , Animais , Avena/química , Modelos Animais de Doenças , Glicogênio Sintase Quinase 3 beta/metabolismo , Hipocampo/metabolismo , Potenciação de Longa Duração/fisiologia , Masculino , Camundongos , Camundongos Transgênicos , Plasticidade Neuronal , Extratos Vegetais/farmacologiaRESUMO
Recent advances in high-throughput sequencing technologies and data science have facilitated the development of precision medicine to treat cancer patients. Synthetic lethality is one of the core methodologies employed in precision cancer medicine. Synthetic lethality describes the phenomenon of the interplay between two genes in which deficiency of a single gene does not abolish cell viability but combined deficiency of two genes leads to cell death. In cancer treatment, synthetic lethality is leveraged to exploit the dependency of cancer cells on a pathway that is essential for cell survival when a tumor suppressor is mutated. This approach enables pharmacological targeting of mutant tumor suppressors that are theoretically undruggable. Successful clinical introduction of BRCA-PARP synthetic lethality in cancer treatment led to additional discoveries of novel synthetic lethal partners of other tumor suppressors, including p53, PTEN, and RB1, using high-throughput screening. Recent work has highlighted aurora kinase A (AURKA) as a synthetic lethal partner of multiple tumor suppressors. AURKA is a serine/threonine kinase involved in a number of central biological processes, such as the G2/M transition, mitotic spindle assembly, and DNA replication. This review introduces synthetic lethal interactions between AURKA and its tumor suppressor partners and discusses the potential of AURKA inhibitors in precision cancer medicine.
Assuntos
Aurora Quinase A/genética , Aurora Quinase A/metabolismo , Biomarcadores Tumorais , Neoplasias/etiologia , Neoplasias/metabolismo , Inibidores de Proteínas Quinases/farmacologia , Mutações Sintéticas Letais , Animais , Ensaios Clínicos como Assunto , Suscetibilidade a Doenças , Desenvolvimento de Medicamentos , Avaliação Pré-Clínica de Medicamentos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Terapia de Alvo Molecular , Neoplasias/tratamento farmacológico , Neoplasias/patologia , Medicina de Precisão , Inibidores de Proteínas Quinases/uso terapêutico , Transdução de Sinais/efeitos dos fármacos , Resultado do TratamentoRESUMO
Nasopharyngeal carcinoma (NPC) is a malignant head and neck cancer type with high morbidity in Southeast Asia, however the pathogenic mechanism of this disease is poorly understood. Using integrative pharmacogenomics, we find that NPC subtypes maintain distinct molecular features, drug responsiveness, and graded radiation sensitivity. The epithelial carcinoma (EC) subtype is characterized by activations of microtubule polymerization and defective mitotic spindle checkpoint related genes, whereas sarcomatoid carcinoma (SC) and mixed sarcomatoid-epithelial carcinoma (MSEC) subtypes exhibit enriched epithelial-mesenchymal transition (EMT) and invasion promoting genes, which are well correlated with their morphological features. Furthermore, patient-derived organoid (PDO)-based drug test identifies potential subtype-specific treatment regimens, in that SC and MSEC subtypes are sensitive to microtubule inhibitors, whereas EC subtype is more responsive to EGFR inhibitors, which is synergistically enhanced by combining with radiotherapy. Through combinational chemoradiotherapy (CRT) screening, effective CRT regimens are also suggested for patients showing less sensitivity to radiation. Altogether, our study provides an example of applying integrative pharmacogenomics to establish a personalized precision oncology for NPC subtype-guided therapies.
Assuntos
Carcinoma Nasofaríngeo/tratamento farmacológico , Carcinoma Nasofaríngeo/genética , Neoplasias Nasofaríngeas/tratamento farmacológico , Neoplasias Nasofaríngeas/genética , Farmacogenética/métodos , Avaliação Pré-Clínica de Medicamentos/métodos , Transição Epitelial-Mesenquimal , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Técnicas de Inativação de Genes , Humanos , Pessoa de Meia-Idade , Carcinoma Nasofaríngeo/patologia , Neoplasias Nasofaríngeas/patologia , Medicina de Precisão , Transcriptoma , Sequenciamento do ExomaRESUMO
This study examined the effects of Mentha arvensis (MA) and Geranium thunbergii (GT) extracts in drinking water on the production performance, egg quality, cholesterol content of egg yolk, proximate composition, and sensory qualities of egg and immunity parameters in laying hens. Ninety-six 28-week-old Hy-Line Brown layers were randomly divided into four dietary treatments for 16 weeks. The dietary treatments were (1) control, (2) T1 (0.01% 1 MA:1 GT), (3) T2 (0.05% 1 MA:1 GT), and (4) T3 (0.1% 1 MA:1 GT). Egg production increased significantly with increasing levels of MA and GT. The egg weight was increased in T2, and the feed intake was highest in T2 and T3 (p < 0.05). The Haugh unit and egg shape index were significantly better in T3 and the control than with other treatments (p < 0.05). The content of yolk cholesterol was significantly lower (p < 0.05) in T2 and T3. On the other hand, there were no significant differences in the egg proximate composition. A significant increase in the serum interleukin 6 (IL-6), tumor necrosis factor (TNFα) and immunoglobulins (IgG and IgA) concentration was observed in the birds fed plant extracts when compared to the control. On average, T2 and T3 showed significantly lower (p < 0.05) concentrations of NH3 gas from the feces as compared to the control. This study suggests that MA and GT supplementation could improve the laying performance, egg quality, and immunity, and decrease the egg yolk cholesterol content in a dose-dependent manner.
RESUMO
Capsidiol, is an anti-fungal phytoalexin produced by plants of Solanaceae. Capsidiol was examined in cultures of primary splenocytes (SPLCs) isolated from healthy C57BL/6 mice and from those with induced experimental autoimmune encephalomyelitis (EAE) as a mouse model for autoimmune neurodegenerative multiple sclerosis (MS). We also examined the impact of capsidiol in IFN-γ-stimulated mouse BV2 microglial cells. Capsidiol resulted in a significant reduction in the anti-CD3/CD28 (αCD3/CD28)-induced IFN-γ+ CD4+ (Th1) and IFN-γ+ CD8+ (Tc1) populations as well as in the production of cytokines (IFN-γ, IL-17A, IL-6, IL-2, TNF-α, and IP-10). Specifically, the CD4+ and CD8+ populations (T-bet+ IFN-γ- , T-bet+ IFN-γ+ , and T-bet- IFN-γ+ ) and cytokine production mediated by Th1/Tc1 polarization were diminished by 25 µM capsidiol. MOG35-55 restimulation of SPLCs from EAE mice resulted in an increase in antigen-specific T cells, including Th1, IL-17A+ CD4+ (Th17), and IL-17A+ CD8+ (Tc17) populations. By contrast, capsidiol resulted in a decrease in the proportions of Th17 and Tc17 cells; MOG35-55 -specific cytokine production was also diminished by capsidiol. Capsidiol treatment resulted in diminished levels of IFN-γ-induced nitric oxide and IL-6; expression of iNOS and COX-2 were suppressed by 50 µM capsidiol in IFN-γ-stimulated BV2 cells. This is the first report of capsidiol-mediated immunomodulatory and antineuroinflammatory activities that may serve to prevent neurodegeneration.
Assuntos
Capsicum/química , Inflamação/tratamento farmacológico , Esclerose Múltipla/tratamento farmacológico , Sesquiterpenos/uso terapêutico , Baço/efeitos dos fármacos , Animais , Modelos Animais de Doenças , Feminino , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Sesquiterpenos/farmacologia , Baço/metabolismoRESUMO
This feeding trial was conducted to investigate the effects of Mentha arvensis (MA) and Geranium thunbergii (GT) in drinking water on physicochemical attributes, sensory qualities, proximate analysis and oxidative stability of broiler leg meat. One hundred and twenty broiler chicks were assigned to 1 of 4 dietary treatments for 5 weeks. The dietary treatments were 1) control, 2) T1 (0.1% 1 MA:1 GT), 3) T2 (0.1% 1 MA:4 GT), 4) T3 (0.1% 4 MA: 1 GT). The water holding capacity and cooking loss were improved (p < 0.05) in T2 and T3. The flavor, texture and acceptability of leg meat by consumers were significantly increased in T2 relative to the control (p < 0.05). The crude protein content was increased in T3 while the crude fat decreased in T2 (p < 0.05). Moreover, broilers supplemented with plant extracts had the lowest leg meat TBARS (thiobarbituric acid reactive substances) values after 2 weeks of storage as compared with the control. Total phenolic contents and 1-1-diphenyl 2 picrylhydrazyl (DPPH) activity were also better in the T2 group (p < 0.05) compared with the control, whereas 2,2-Azinobis-3 ethytlbenzothiazoline-6-sulfonic acid (ABTS+) remained unaffected. Overall, these results demonstrate that broiler drinking water with the inclusion of plant extract combination can be used to enhance the oxidative stability, shelf life and quality characteristics of broiler leg meat without compromising the growth performance.
RESUMO
S100A8 and S100A9 are important proteins in the pathogenesis of allergy. Asthma is an allergic lung disease, characterized by bronchial inflammation due to leukocytes, bronchoconstriction, and allergen-specific IgE. In this study, we examined the role of S100A8 and S100A9 in the interaction of cytokine release from bronchial epithelial cells, with constitutive apoptosis of neutrophils. S100A8 and S100A9 induce increased secretion of neutrophil survival cytokines such as MCP-1, IL-6 and IL-8. This secretion is suppressed by TLR4 inhibitor), LY294002, AKT inhibitor, PD98059, SB202190, SP600125, and BAY-11-7085. S100A8 and S100A9 also induce the phosphorylation of AKT, ERK, p38 MAPK and JNK, and activation of NF-κB, which were blocked after exposure to TLR4i, LY294002, AKTi, PD98059, SB202190 or SP600125. Furthermore, supernatants collected from bronchial epithelial cells after S100A8 and S100A9 stimulation suppressed the apoptosis of normal and asthmatic neutrophils. These inhibitory mechanisms are involved in suppression of caspase 9 and caspase 3 activation, and BAX expression. The degradation of MCL-1 and BCL-2 was also blocked by S100A8 and S100A9 stimulation. Essentially, neutrophil apoptosis was blocked by co-culture of normal and asthmatic neutrophils with BEAS-2B cells in the presence of S100A8 and S100A9. These findings will enable elucidation of asthma pathogenesis.
Assuntos
Asma/metabolismo , Calgranulina A/uso terapêutico , Calgranulina B/farmacologia , Apoptose/efeitos dos fármacos , Western Blotting , Caspase 3/metabolismo , Caspase 9/metabolismo , Linhagem Celular , Citocinas/metabolismo , Ensaio de Imunoadsorção Enzimática , Eosinófilos/efeitos dos fármacos , Eosinófilos/metabolismo , Humanos , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Receptor 4 Toll-Like/metabolismoRESUMO
Glycyrrhizae Radix is widely used as herbal medicine and is effective against inflammation, various cancers, and digestive disorders. We aimed to develop a sensitive and simultaneous analytical method for detecting glycyrrhizin, isoliquiritigenin, liquiritigenin, and liquiritin, the four marker components of Glycyrrhizae Radix extract (GRE), in rat plasma using liquid chromatography-tandem mass spectrometry and to apply this analytical method to pharmacokinetic studies. Retention times for glycyrrhizin, isoliquiritigenin, liquiritigenin, and liquiritin were 7.8 min, 4.1 min, 3.1 min, and 2.0 min, respectively, suggesting that the four analytes were well separated without any interfering peaks around the peak elution time. The lower limit of quantitation was 2 ng/mL for glycyrrhizin and 0.2 ng/mL for isoliquiritigenin, liquiritigenin, and liquiritin; the inter- and intra-day accuracy, precision, and stability were less than 15%. Plasma concentrations of glycyrrhizin, isoliquiritigenin, liquiritigenin, and liquiritin were quantified for 24 h after a single oral administration of 1 g/kg GRE to four rats. Among the four components, plasma concentration of glycyrrhizin was the highest and exhibited a long half-life (23.1 ± 15.5 h). Interestingly, plasma concentrations of isoliquiritigenin and liquiritigenin were restored to the initial concentration at 4-10 h after the GRE administration, as evidenced by liquiritin biotransformation into isoliquiritigenin and liquiritigenin, catalyzed by fecal lysate and gut wall enzymes. In conclusion, our analytical method developed for detecting glycyrrhizin, isoliquiritigenin, liquiritigenin, and liquiritin could be successfully applied to investigate their pharmacokinetic properties in rats and would be useful for conducting further studies on the efficacy, toxicity, and biopharmaceutics of GREs and their marker components.
Assuntos
Chalconas/sangue , Flavanonas/sangue , Glucosídeos/sangue , Ácido Glicirrízico/sangue , Administração Oral , Animais , Chalconas/farmacocinética , Cromatografia Líquida , Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/farmacocinética , Flavanonas/farmacocinética , Glucosídeos/farmacocinética , Ácido Glicirrízico/farmacocinética , Masculino , Extratos Vegetais/sangue , Extratos Vegetais/farmacocinética , Controle de Qualidade , Ratos , Ratos Sprague-Dawley , Espectrometria de Massas em TandemRESUMO
BACKGROUND: Multiple sclerosis (MS) is an autoimmune disorder resulting in paralysis, and the responses of reactive T cells against self-antigens are hallmarks. Glycyrrhizae Radix (GR) has been used for detoxification and reducing inflammation. However, very few reports have described the effects of GR on MS. PURPOSE: The immunomodulatory effects of GR extract on autoimmune responses were evaluated through in vitro, ex vivo, and in vivo assays using primary mouse splenocytes (SPLC), mouse microglia BV2 cell line, and a mouse model of experimental autoimmune encephalomyelitis (EAE). STUDY DESIGN: Ethanol extract of GR was used in vitro with primary SPLC in the condition of anti-CD3/CD28 stimulation and interferon (IFN)-γ-producing CD4+ (TH1)/CD8+ (TC1) polarization as well as IFN-γ-stimulated BV2 cells. For EAE induction, female C57BL/6 mice were immunized with 200⯵g of myelin oligodendrocyte glycoprotein (MOG)35-55 without pertussis toxin. EAE SPLC (ex vivo) and EAE mice (in vivo) were treated with GR extract to evaluate the changes in antigen-specific responses. SPLC media containing antigen-specific responses were used to stimulate BV2 cells. RESULTS: GR extract effectively modulated the responses of reactive splenic T cells through the reduction in IFN-γ+ T cell populations, the expressions of cell adhesion molecules (CAMs), and secretions of cytokines containing IFN-γ and a chemokine IFN-γ-induced protein 10 (IP-10) in vitro. In addition, GR extract significantly decreased nitric oxide production and secretion of tumor necrosis factor (TNF)-α and IP-10 in IFN-γ-stimulated BV2 cells. The antigen-specific TH1 and TC1 populations were decreased following administration of 100â¯mg/kg of GR extract, whereas CD8+IL-17A+ (TC17) population was increased on day 36 after EAE induction. Moreover, IFN-γ, which showed the highest secretion among examined cytokines, and IP-10 decreased on day 36. SPLC media derived from 100â¯mg/kg GR extract-administered EAE mice revealed the ameliorative effects on BV2 cell stimulation. CONCLUSION: This is the first report on the immunomodulatory effects of GR extract on antigen-specific SPLC responses in EAE. These results could be helpful for the discovery of drug candidates for MS by focusing on IFN-γ-related autoimmune responses.
Assuntos
Encefalomielite Autoimune Experimental/tratamento farmacológico , Glycyrrhiza/química , Extratos Vegetais/farmacologia , Baço/imunologia , Animais , Células Cultivadas , Citocinas/metabolismo , Encefalomielite Autoimune Experimental/imunologia , Encefalomielite Autoimune Experimental/metabolismo , Etanol/química , Feminino , Interferon gama/metabolismo , Interleucina-17 , Camundongos Endogâmicos C57BL , Microglia/efeitos dos fármacos , Glicoproteína Mielina-Oligodendrócito/imunologia , Óxido Nítrico/metabolismo , Fragmentos de Peptídeos/imunologia , Extratos Vegetais/química , Baço/citologia , Baço/efeitos dos fármacos , Células Th1/efeitos dos fármacos , Células Th1/imunologiaRESUMO
The methanol extract of Abronia nana suspension cultures were subjected to column chromatography to identify potential inhibitors of ß-secretase, which is a major factor in Alzheimer's disease development. Two new C-methylisoflavones boeravinone T (1) and U (4) were isolated with three knowns boeravinone B (2), J (3) and X (5). The half-maximal inhibitory concentration (IC50) values of compounds 1-5 were 18.29, 8.57, 7.87, 12.02 and 5.30 µM, respectively. The most potent 5, non-competitively inhibited ß-secretase [inhibition constant (Ki) = 3.79 µM]. Compounds 1-5 did not inhibit other proteases such as chymotrypsin, trypsin and elastase at concentrations up to 1 mM, indicating that they were relatively specific inhibitors of ß-secretase. A free hydroxyl group at C-3 position of the C-methylisoflavone skeleton appeared to be responsible for the stronger inhibitory activity against ß-secretase.
Assuntos
Secretases da Proteína Precursora do Amiloide/antagonistas & inibidores , Nyctaginaceae/química , Doença de Alzheimer/etiologia , Benzopiranos/isolamento & purificação , Benzopiranos/farmacologia , Flavonoides/isolamento & purificação , Flavonoides/farmacologia , Humanos , Concentração Inibidora 50 , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Relação Estrutura-Atividade , Especificidade por SubstratoRESUMO
ARID1A, a component of the SWI/SNF chromatin remodeling complex, is a tumor suppressor with a high frequency of inactivating mutations in many cancers. Therefore, ARID1A deficiency has been exploited therapeutically for treating cancer. Here we show that ARID1A has a synthetic lethal interaction with aurora kinase A (AURKA) in colorectal cancer (CRC) cells. Pharmacological and genetic perturbations of AURKA selectively inhibit the growth of ARID1A-deficient CRC cells. Mechanistically, ARID1A occupies the AURKA gene promoter and negatively regulates its transcription. Cells lacking ARID1A show enhanced AURKA transcription, which leads to the persistent activation of CDC25C, a key protein for G2/M transition and mitotic entry. Inhibiting AURKA activity in ARID1A-deficient cells significantly increases G2/M arrest and induces cellular multinucleation and apoptosis. This study shows a novel synthetic lethality interaction between ARID1A and AURKA and indicates that pharmacologically inhibiting the AURKA-CDC25C axis represents a novel strategy for treating CRC with ARID1A loss-of-function mutations.
Assuntos
Aurora Quinase A/metabolismo , Neoplasias Colorretais/genética , Proteínas Nucleares/deficiência , Transdução de Sinais , Mutações Sintéticas Letais/genética , Fatores de Transcrição/deficiência , Fosfatases cdc25/metabolismo , Animais , Apoptose , Aurora Quinase A/antagonistas & inibidores , Aurora Quinase A/genética , Neoplasias Colorretais/patologia , Proteínas de Ligação a DNA , Avaliação Pré-Clínica de Medicamentos , Feminino , Fase G2 , Técnicas de Inativação de Genes , Humanos , Camundongos Endogâmicos BALB C , Camundongos Nus , Mitose , Proteínas Nucleares/metabolismo , Fatores de Transcrição/metabolismo , Transcrição GênicaRESUMO
Atopic dermatitis (AD) is a common chronic inflammatory skin disorder afflicting from infancy to adults with itching, scratching, and lichenification. We aimed to investigate the effects of esculetin from Fraxinus rhynchophylla on atopic skin inflammation. For induction of atopic skin inflammation, we exposed the ears of female BALB/c mice to house dust mite (Dermatophagoides farinae extract, DFE) and 2,4-dinitrochlorobenzene (DNCB) for 4â¯weeks. Oral administration of esculetin reduced the symptoms of DFE/DNCB-induced atopic skin inflammation, which were evaluated based on ear swelling and number of scratch bouts. The immunoglobulin (Ig) E, IgG2a, and histamine levels in serum were decreased and inflammatory cell infiltration in skin tissue was reduced by the esculetin. It suppressed production of Th1, Th2 and Th17-related cytokines such as tumor necrosis factor (TNF)-α, interferon (IFN)-γ, interleukin (IL)-4, IL-13, IL-31 and IL-17 in the ear tissue. Furthermore, we investigated the effects of esculetin on activated keratinocytes, which are representative cells used for studying the pathogenesis of acute and chronic atopic skin inflammation. As results, esculetin suppressed gene expression of Th1, Th2 and Th17 cytokines and the activation of nuclear factor-κB and signal transducer and activator of transcription 1 in TNF-α/IFN-γ-stimulated keratinocytes. Taken together, these results imply that esculetin attenuated atopic skin inflammation, suggesting that esculetin could be a potential therapeutic candidate for the treatment of AD.
Assuntos
Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Citocinas/antagonistas & inibidores , Dermatite Alérgica de Contato/tratamento farmacológico , Umbeliferonas/farmacologia , Umbeliferonas/uso terapêutico , Animais , Antígenos de Dermatophagoides , Linhagem Celular , Citocinas/genética , Citocinas/imunologia , Dermatite Alérgica de Contato/imunologia , Dermatite Alérgica de Contato/metabolismo , Dinitroclorobenzeno , Feminino , Fraxinus , Humanos , Queratinócitos/efeitos dos fármacos , Queratinócitos/metabolismo , Camundongos Endogâmicos BALB C , NF-kappa B/metabolismo , Fator de Transcrição STAT1/metabolismo , Pele/efeitos dos fármacos , Pele/imunologiaRESUMO
Enzyme fermentation is a type of food processing technique generally used to improve the biological activities of food and herbal medicines. In this study, a Syzygii Flos (clove) extract was fermented using laccase derived from Trametes versicolor (LTV). The fermented clove extract showed greater neuroprotective effects against glutamate toxicity on HT22 than the non-fermented extract did. HPLC analysis revealed that the eugenol (1) and dehydrodieugenol (2) contents had decreased and increased, respectively, after fermentation. The content of 2 peaked at 1 h after fermentation to 103.50 ± 8.20 mg/gex (not detected at zero time), while that of 1 decreased to 79.54 ± 4.77 mg/gex (185.41 ± 10.16 mg/gex at zero time). Compound 2 demonstrated promising HT22 neuroprotective properties with inhibition of Ca2+ influx, the overproduction of intracellular reactive oxygen species, and lipid peroxidation. In addition, LTV showed the best fermentation efficacy compared with laccases derived from Pleurotus ostreatus and Rhus vernicifera.
Assuntos
Eugenol/análogos & derivados , Fermentação , Ácido Glutâmico/toxicidade , Lacase/metabolismo , Lignanas/metabolismo , Lignanas/farmacologia , Extratos Vegetais/metabolismo , Extratos Vegetais/farmacologia , Syzygium/química , Animais , Morte Celular/efeitos dos fármacos , Linhagem Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Eugenol/química , Eugenol/metabolismo , Eugenol/farmacologia , Proteínas Fúngicas/metabolismo , Lignanas/química , Peroxidação de Lipídeos , Camundongos , Plantas Medicinais , Pleurotus/enzimologia , Pleurotus/metabolismo , Ratos , Espécies Reativas de Oxigênio , República da Coreia , Rhus/enzimologia , Rhus/metabolismo , Trametes/enzimologia , Trametes/metabolismoRESUMO
Herbal medicines were subjected to enzyme reaction by using a commercial glycosidase AMG-300L, and were evaluated for enhancement of their antioxidative activities. The methanolic extract of Gentianae Scabrae Radix (GSR) showed the most dramatic changes after enzyme reaction, as seen in the high-performance liquid chromatography profiles and an increase in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging effect. Trifloroside (1, TF) was identified as being significantly decreased by enzyme reaction, whereas deglucosyltrifloroside (2, DTF) increased. The optimal reaction time to induce DTF was 24 h at 30°C. The content increased from 1.00 ± 0.29 mg/g of extract (gex) to 2.80 ± 0.85 mg/gex after 24 h of enzyme reaction. DTF showed better antioxidative effect than TF in the DPPH, Trolox equivalent antioxidant capacity, and reactive oxygen species (ROS) in HT22 cell assays. In addition, when HT22 cells were stressed by 5 mM glutamate, 50 µM of DTF significantly inhibited the glutamate-induced lactate dehydrogenase leakage, Ca2+ influx, lipid peroxidation, and intracellular ROS production. These data demonstrated that the enzyme-treated GSR and its increased level of antioxidant DTF could be useful as a starting point in the discovery of functional foods to prevent various oxidative stresses, especially neurodegenerative disorders.
Assuntos
Antioxidantes/química , Antioxidantes/farmacologia , Gentianaceae/química , Glucosídeos/química , Glucosídeos/farmacologia , Glicosídeo Hidrolases/química , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Aspergillus niger/enzimologia , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Proteínas Fúngicas/química , Humanos , Peroxidação de Lipídeos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Plantas Medicinais/química , Espécies Reativas de Oxigênio/metabolismoRESUMO
Viscum album var. coloratum (Korean mistletoe; KM) is an herbal medicine that is used worldwide for the treatment of various immunological disorders and cancers. KM extract showed enhanced anti-oxidative effects in 2,2-diphenyl-1-picrylhydrazyl, Trolox equivalent antioxidant capacity, and 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate acetyl ester assays after being fermented with a crude enzyme extract from a soybean paste fungus, Aspergillus kawachii. High-performance liquid chromatography analysis showed four increased peaks in enzyme treated KM. The increased peaks were isolated and identified as caffeic acid (1), hesperetin (2), syringaldehyde (3), and lyoniresinol (4). Among the four compounds, only 1 and 4 showed strong anti-oxidative activity. Therefore, the fermentation increased the contents of 1 and 4, which consequently increased the anti-oxidative activity of KM.
Assuntos
Anisóis/química , Antioxidantes/farmacocinética , Ácidos Cafeicos/química , Fermentação , Erva-de-Passarinho/química , Naftalenos/química , Animais , Anisóis/isolamento & purificação , Antioxidantes/química , Antioxidantes/isolamento & purificação , Aspergillus/metabolismo , Benzaldeídos/química , Benzaldeídos/isolamento & purificação , Ácidos Cafeicos/isolamento & purificação , Linhagem Celular , Cromatografia Líquida de Alta Pressão , Sequestradores de Radicais Livres/análise , Ácido Glutâmico/metabolismo , Medicina Herbária , Hesperidina/química , Hesperidina/isolamento & purificação , Camundongos , Naftalenos/isolamento & purificação , Fármacos Neuroprotetores , Extratos Vegetais/química , Extratos Vegetais/farmacologia , SolventesRESUMO
The ubiquitous nuclear protein, high mobility group box-1 (HMGB1) functions as a late mediator of sepsis. A new rarely occurring C-methylrotenoid, named boeravinone X (comp 1), was isolated and identified from Abronia nana suspension cultures during our continuous works on the discovery of anti-septic natural products. Here, we investigated the antiseptic effects and underlying mechanisms of comp 1 against HMGB1-mediated septic responses. According to the results, comp 1 effectively inhibited lipopolysaccharide (LPS)-induced release of HMGB1, and suppressed HMGB1-mediated septic responses, such as hyperpermeability, adhesion and migration of leukocytes, and expression of cell adhesion molecules. And, comp 1 suppressed the production of tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6), and the activation of nuclear factor-κB (NF-κB) and extracellular signal-regulated kinases 1 and 2 (ERK1/2) by HMGB1. Collectively, these results indicate that comp 1 could be potential therapeutic agents for the treatment of various severe vascular inflammatory diseases via inhibition of the HMGB1 signaling pathway.
Assuntos
Anti-Inflamatórios/farmacologia , Benzopiranos/farmacologia , Flavonoides/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Proteína HMGB1/genética , Nyctaginaceae/química , Sepse/tratamento farmacológico , Animais , Anti-Inflamatórios/isolamento & purificação , Benzopiranos/isolamento & purificação , Movimento Celular/efeitos dos fármacos , Movimento Celular/imunologia , Flavonoides/isolamento & purificação , Proteína HMGB1/antagonistas & inibidores , Proteína HMGB1/imunologia , Proteína HMGB1/metabolismo , Células Endoteliais da Veia Umbilical Humana , Humanos , Interleucina-6/antagonistas & inibidores , Interleucina-6/genética , Interleucina-6/imunologia , Leucócitos/efeitos dos fármacos , Leucócitos/imunologia , Leucócitos/patologia , Lipopolissacarídeos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteína Quinase 1 Ativada por Mitógeno/antagonistas & inibidores , Proteína Quinase 1 Ativada por Mitógeno/genética , Proteína Quinase 1 Ativada por Mitógeno/imunologia , Proteína Quinase 3 Ativada por Mitógeno/antagonistas & inibidores , Proteína Quinase 3 Ativada por Mitógeno/genética , Proteína Quinase 3 Ativada por Mitógeno/imunologia , NF-kappa B/antagonistas & inibidores , NF-kappa B/genética , NF-kappa B/imunologia , Extratos Vegetais/química , Sepse/induzido quimicamente , Sepse/genética , Sepse/patologia , Transdução de Sinais , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Beyond its role in the activation of protein C, the endothelial cell protein C receptor (EPCR) plays an important role in the cytoprotective pathway. EPCR can be shed from the cell surface, which is mediated by tumor necrosis factor-α converting enzyme (TACE). Polyozellin, a major constituent of a Korea edible mushroom Polyozellus multiplex, has been known to exhibit the biological activities such as anti-oxidative and anti-inflammatory effects. However, little is known about the effects of polyozellin on EPCR shedding. We investigated this issue by monitoring the effects of polyozellin on phorbol-12-myristate 13-acetate (PMA)-, tumor necrosis factor (TNF)-α-, interleukin (IL)-1ß-induced EPCR shedding in human umbilical vein endothelial cells (HUVECs), and cecal ligation and puncture (CLP)-mediated EPCR shedding in mice and underlying mechanism. Data demonstrate that polyozellin induced potent inhibition of PMA-, TNF-α-, IL-1ß- (in HUVECs), and CLP-induced EPCR shedding (in mice) via inhibition of phosphorylation of mitogen-activated protein kinases (MAPKs) such as p38, janus kinase (JNK), and extracellular signal-regulated kinase (ERK) 1/2. Polyozellin also inhibited the expression and activity of PMA-induced TACE in HUVECs suggesting that p38, ERK1/2, and JNK could be the molecular targets of POZ. These results demonstrate the potential of polyozellin as an anti-EPCR shedding reagent against PMA-mediated and CLP-mediated EPCR shedding.
Assuntos
Proteínas ADAM/antagonistas & inibidores , Agaricales/química , Antígenos CD/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/antagonistas & inibidores , Furanos/química , Receptores de Superfície Celular/metabolismo , Proteína ADAM17 , Animais , Modelos Animais de Doenças , Receptor de Proteína C Endotelial , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Humanos , Interleucina-1beta/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Sepse/metabolismo , Acetato de Tetradecanoilforbol/farmacologia , Fator de Necrose Tumoral alfa/farmacologiaRESUMO
It is thought that the neuronal cell loss caused by oxidative stress is the primary mechanism underlying the pathogenesis of several neurodegenerative disorders. Glutamate is an endogenous neurotransmitter, but at high concentrations it can act as a neurotoxicant by increasing the intracellular levels of reactive oxygen species (ROS). Therefore, the development of factors that can attenuate glutamate-induced oxidative stress in neuronal cells is a good strategy by which new drugs could be discovered that may treat or prevent neurodegenerative diseases. Here, the neuroprotective effects of kaempferol (KF) isolated from the stems of butterbur (Petasites japonicus) were examined in glutamate-treated hippocampal neuronal cells (HT22). The administration of KF (25 µM) resulted in a significant increase in cell viability (105.18 ± 7.48%) compared with the control (100.00 ± 3.05%), while glutamate (5 mM) reduced cell viability by 39.94 ± 1.61%. The glutamate-induced calcium (Ca(2+)) influx (1.93 ± 0.08-fold) was significantly reduced by 0.89 ± 0.02-fold following the administration of 25 µM KF. Additionally, when HT22 cells were stressed with excessive glutamate, there was a 3.70 ± 0.01-fold increase in intracellular ROS generation, even though this was effectively attenuated by KF (25 µM, 0.72 ± 0.01-fold). The protective effects of KF in HT22 cells were later confirmed using a lactate dehydrogenase (LDH) assay and a FITC-annexin V/propidium iodide double staining procedure. These findings also revealed that the neuroprotective effects of KF are a result of the regulation of the expression levels of proteins, such as Bcl-2, Bid, apoptosis-inducing factor (AIF), and mitogen-activated protein kinase (MAPK). This is the first report to investigate the neuroprotective influence of KF in glutamate-treated HT22 cells. These data demonstrate that KF may be a useful candidate for pharmacological therapies that can prevent and treat neurodegenerative diseases such as Alzheimer's disease (AD).
Assuntos
Ácido Glutâmico/efeitos adversos , Hipocampo/citologia , Quempferóis/farmacologia , Neurônios/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Animais , Anexina A5/metabolismo , Apoptose/efeitos dos fármacos , Fator de Indução de Apoptose/genética , Fator de Indução de Apoptose/metabolismo , Proteína Agonista de Morte Celular de Domínio Interatuante com BH3/genética , Proteína Agonista de Morte Celular de Domínio Interatuante com BH3/metabolismo , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Fluoresceína-5-Isotiocianato/análogos & derivados , Fluoresceína-5-Isotiocianato/metabolismo , Ácido Glutâmico/administração & dosagem , Camundongos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Fármacos Neuroprotetores/farmacologia , Petasites/química , Fosforilação , Extratos Vegetais/farmacologia , Propídio/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismoRESUMO
This study was performed to compare the dietary food and nutrient intakes according to supplement use in pregnant and lactating women in Seoul. The subjects were composed of 201 pregnant and 104 lactating women, and their dietary food intake was assessed using the 24-h recall method. General information on demographic and socioeconomic factors, as well as health-related behaviors, including the use of dietary supplements, were collected. About 88% and 60% of the pregnant and lactating women took dietary supplements, respectively. The proportion of dietary supplements used was higher in pregnant women with a higher level of education. After adjusting for potential confounders, among the pregnant women, supplement users were found to consume 45% more vegetables, and those among the lactating women were found to consume 96% more beans and 58% more vegetables. The intakes of dietary fiber and ß-carotene among supplement users were higher than those of non-users, by 23% and 39%, respectively. Among pregnant women, the proportion of women with an intake of vitamin C (from diet alone) below the estimated average requirements (EAR) was lower among supplement users [users (44%) vs. non-users (68%)], and the proportion of lactating women with intakes of iron (from diet alone) below the EAR was lower among supplement users [usesr (17%) vs. non-users (38%)]. These results suggest that among pregnant and lactating women, those who do not use dietary supplements tend to have a lower intake of healthy foods, such as beans and vegetables, as well as a lower intake of dietary fiber and ß-carotene, which are abundant in these foods, and non-users are more likely than users to have inadequate intake of micro-nutrient such as vitamin C and iron.