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To determine the role of the cystine/glutamate antiporter on retinal structure and function, retinas of C57Bl/6J wild-type and xCT knockout mice, lacking the xCT subunit of the cystine/glutamate antiporter were examined from 6 weeks to 12 months of age. Fundoscopy, optical coherence tomography (OCT), and whole mount retinal autofluorescence imaging were used to visualise age-related retinal spots. Glial fibrillary acidic protein (GFAP) immunolabelling was used to assess retinal stress. Retinal function was evaluated using full-field and focal electroretinograms. Examinations revealed retinal spots in both wild-type and xCT knockout mice with the number of spots greater at 9 months in the knockout compared to wild-type. OCT confirmed these discrete spots were located at the retinal pigment epithelium (RPE)-photoreceptor junction and did not label with drusen markers. Whole mount lambda scans of the 9 month xCT knockout retinas revealed that the photoreceptor autofluorescence matched the spots, suggesting these spots were retinal debris. GFAP labelling was increased in knockout retinas compared to wild-type indicative of retinal stress, and the discrete spots were associated with migration of microglia/macrophages to the RPE-retina intersection. OCT revealed that the superior retina was thinner at 9 months in knockout compared to wild-type mice due to changes to the outer nuclear and photoreceptor layers. While global retinal function was not affected by loss of xCT, focal changes in retinal function were detected in areas where spots were present. Tother these results suggest that the xCT KO mice exhibit features of accelerated ageing and suggests that this mouse model may be useful for studying the underlying cellular pathways in retinal ageing.
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Cistina , Ácido Glutâmico , Camundongos , Animais , Cistina/metabolismo , Camundongos Knockout , Ácido Glutâmico/metabolismo , Retina/metabolismo , Epitélio Pigmentado da Retina/metabolismo , Camundongos Endogâmicos C57BLRESUMO
The production of ε-poly-L-lysine (ε-PL) from cassava bagasse hydrolysate (CBH) by Streptomyces albulus US3-18 was investigated in this study. With 30 g/L glucose from CBH, 1.30 g/L ε-PL and 10.68 g/L biomass were obtained in shake flask fermentation. Interestingly, the two values were increased by 14.0% and 21.5%, respectively, compared to the control (1.14 g/L and 8.79 g/L). Simultaneously, the activities of four key enzymes of ε-PL synthesis during CBH fermentation were enhanced to varying degrees. In batch fermentation of 5-L bioreactor, 3.39 g/L ε-PL and 10.17 g/L DCW were harvested with 40 g/L glucose from CBH. The combination of fed-batch fermentation with two-stage pH strategy significantly increased ε-PL titer and biomass to 37.41 g/L and 41.0 g/L, respectively. Moreover, eleven volatile components were detected in CBH by GC-MS, and 6-pentyl-α-pyrone (6PP) was first identified as the most abundant volatile ingredient. The results in CBH fermentation demonstrated that S. albulus US3-18 exhibited high tolerance to these volatile byproducts. Using ICP-MS, the calcium concentration in CBH was determined as 195.0 mg/(kg hydrolyzate), and cobalt, copper, lead, chromium, mercury and arsenic were not detected. By adding 0.05 g/L CaCl2 to M3G medium, ε-PL yield was improved by 28.0%, indicating calcium was one of the factors for the enhanced ε-PL production. The study provides a reference for the efficient production of ε-PL from low-cost agricultural residues.
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Manihot , Polilisina , Cálcio , Carbono , Celulose , Fermentação , Glucose , StreptomycesRESUMO
ε-Poly-l-lysine (ε-PL) is used as a natural food preservative which consists of l-lysine units connected. However, due to the expensive culture medium, the production cost of ε-PL remains high. In this study, cheap raw materials cassava starch (CS) and fish meal (FM) were employed by S. albulus FQC-24 for ε-PL production. In the single factor experiment, the maximum ε-PL production reached 0.97 g/L at 60 g/L CS and 15 g/L FM. The results of screening experiments by Plackett-Burman design showed that three main components affecting ε-PL production were CS, FM, and (NH4)2SO4. And the standardized effects of CS, FM, and (NH4)2SO4 were 0.13, -0.22, and -0.2, respectively. The optimum fermentation medium developed by response surface methodology for ε-PL production contained (g/L) CS, 67.56; FM, 14.70 and (NH4)2SO4, 5.41. Under the optimum conditions, the ε-PL production was achieved 1.30 g/L, with 34.02% higher than that before optimization. Moreover, ε-PL productions of batch and fed-batch fermentation in a 7-L fermentor were improved to 2.13 and 17.17 g/L respectively, which increased by 0.64 and 12.2 times compared with the shake flask culture. The results indicated that FM and CS are promising substrates for the efficient production of ε-PL.
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Manihot , Streptomyces , Meios de Cultura/farmacologia , Fermentação , Polilisina/farmacologia , AmidoRESUMO
Vine tea was bio-transformed using Poria cocos by solid-state fermentation in order to improve its taste and quality. Volatile components in vine tea were also identified by GC-MS. The changes of flavonoid, tea polyphenols and polysaccharides in fermented vine tea were evaluated. Flavonoid and polyphenols in vine tea were remained unchanged even after biotransformation, but content of polysaccharides increased to 3.9-fold than that of unfermented vine tea. Antioxidant activity such as DPPH free radical scavenging capacity (SR) was determined that there was a positive correlation between SR and content of polysaccharides in vine tea. Methyl 2-methylvalerate-a new volatile compound was identified and gave the vine tea rich delicate fragrance of fruits. The content of linolenic acid increased from 0.88 to 19.59 %. Biotransformation improved the taste and quality of vine tea.
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This study developed a simple, efficient method for producing racemic ß-phenylalanine acid (BPA) and its derivatives via the enantioselective acylation catalyzed by the penicillin G acylase from Alcaligenes faecalis (Af-PGA). When the reaction was run at 25°C and pH 10 in an aqueous medium containing phenylacetamide and BPA in a molar ratio of 2:1, 8 U/mL enzyme and 0.1 M BPA, the maximum BPA conversion efficiency at 40 min only reached 36.1%, which, however, increased to 42.9% as the pH value and the molar ratio of phenylacetamide to BPA were elevated to 11 and 3:1, respectively. Under the relatively optimum reaction conditions, the maximum conversion efficiencies of BPA derivatives all reached about 50% in a relatively short reaction time (45-90 min). The enantiomeric excess value of product (ee(p)) and enantiomeric excess value of substrate (ee(s)) were all above 98% and 95%, respectively. These results suggest that the method established in this study is practical, effective, and environmentally benign and may be applied to industrial production of enantiomerically pure BPA and its derivatives.
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Acetamidas/química , Alcaligenes faecalis/enzimologia , Penicilina Amidase/química , Fenilalanina/análogos & derivados , Fenilalanina/síntese química , Acilação , Alcaligenes faecalis/genética , Catálise , Ativação Enzimática , Ensaios Enzimáticos/métodos , Estabilidade Enzimática , Escherichia coli/química , Escherichia coli/genética , Penicilina Amidase/genética , Fenilacetatos/química , Fenilalanina/química , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Estereoisomerismo , Fatores de TempoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Centella asiatica (L.) Urb., a potential medicinal plant, is widely used in orient traditional medicine. Its major active constituents include asiaticoside (AS), madecassoside (MS), asiatic acid and madecassic acid. Thereinto, AS is a pentacyclic triterpenoid saponin with a variety of pharmacological effects including antitumor, neuroprotective and wound healing effects. AIM OF THE STUDY: In this review, we summarize the pharmacokinetics, safety and pharmacological properties of AS. MATERIALS AND METHODS: We gathered information about AS from articles published up to 2022 and listed in Google scholar, PubMed, Web of Science, Elsevier, and similar databases. The keywords used in our search included "asiaticoside", "Centella asiatica", "pharmacokinetics", "nerve", "cancer", "skin", etc. RESULTS: AS appeared to degrade through a first-order reaction and had low biotoxicity. However, the pharmacokinetic properties of AS differed according to species. AS is highly blood-brain-barrier permeable without any harmful side effect. It has a variety of pharmacological effects including anti-neural inflammation and anti-cancer properties, as well as protective properties for the skin, cardiovascular system, and pulmonary system. CONCLUSION: This review comprehensively summarized current information regarding the pharmacokinetic and pharmacological properties of AS, and supported the pharmaceutical value of this compound. Future research should focus on improving bioavailability of AS and conducting clinical assessment.
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Centella , Triterpenos , Extratos Vegetais/farmacologia , Triterpenos/farmacologiaRESUMO
Garlic (Allium sativum L.), the underground bulb of the Allium plant in the family Liliaceae, is a common and popular spice that has historically been used to prevent and treat many different diseases such as pain, deafness, diarrhea, tumors, and other healthy problems. Garlic essential oil contains a variety of organosulfur compounds, such as the most representative diallyl disulfides (DADS) and diallyl trisulfides (DATS), which have attracted great interest in medicine, food, and agriculture because of their rich biological activities. This paper reviews the research progress on the composition and bioactivities of garlic essential oil mixtures and the bioactivity of some typical monomeric sulfides in garlic essential oil. The active mechanisms of representative sulfides in garlic essential oil were analyzed, and the applications of garlic essential oil in functional food, food additives, and clinical treatment were discussed. Combined with the current research status, the limitations and development direction of garlic essential oil in the study of molecular mechanism were discussed, which is of great significance to the development of garlic essential oil as a natural and safe alternative medicine for treatment.
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A high-speed counter-current chromatography (HSCCC) method was established for the preparative separation of three sesquiterpenoid lactones from Eupatorium lindleyanum DC. The two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (1:4:2:3, v/v/v/v) was selected. From 540 mg of the n-butanol fraction of Eupatorium lindleyanum DC., 10.8 mg of 3ß-hydroxy-8ß-[4'-hydroxytigloyloxy]-costunolide, 17.9 mg of eupalinolide A and 19.3 mg of eupalinolide B were obtained in a one-step HSCCC separation, with purities of 91.8%, 97.9% and 97.1%, respectively, as determined by HPLC. Their structures were further identified by ESI-MS and ¹H-NMR.
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Medicamentos de Ervas Chinesas/isolamento & purificação , Eupatorium/química , Lactonas/isolamento & purificação , Sesquiterpenos de Germacrano/isolamento & purificação , Sesquiterpenos/isolamento & purificação , Acetatos/química , Butanóis/química , Cromatografia Líquida de Alta Pressão , Distribuição Contracorrente , Medicamentos de Ervas Chinesas/química , Etanol/química , Hexanos/química , Lactonas/química , Metanol/química , Estrutura Molecular , Sesquiterpenos/química , Sesquiterpenos de Germacrano/química , Solventes/química , Espectrometria de Massas por Ionização por Electrospray , Água/químicaRESUMO
Low temperature germination (LTG) is a key agronomic trait in rice (Oryza sativa L.). However, the genetic basis of natural variation for LTG is largely unknown. Here, a genome-wide association study (GWAS) was performed using 276 accessions from the 3,000 Rice Genomes (3K-RG) project with 497 k single nucleotide polymorphisms (SNPs) to uncover potential genes for LTG in rice. In total, 37 quantitative trait loci (QTLs) from the 6th day (D6) to the 10th day (D10) were detected in the full population, overlapping with 12 previously reported QTLs for LTG. One novel QTL, namely qLTG1-2, was found stably on D7 in both 2019 and 2020. Based on two germination-specific transcriptome datasets, 13 seed-expressed genes were isolated within a 200 kb interval of qLTG1-2. Combining with haplotype analysis, a functional uncharacterized gene, LOC_Os01g23580, and a seed germination-associated gene, LOC_Os01g23620 (OsSar1a), as promising candidate genes, both of which were significantly differentially expressed between high and low LTG accessions. Collectively, the candidate genes with favorable alleles may be useful for the future characterization of the LTG mechanism and the improvement of the LTG trait in rice breeding.
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Estudo de Associação Genômica Ampla , Oryza , Oryza/genética , Germinação/genética , Temperatura , Melhoramento VegetalRESUMO
The antioxidant activities of water extract (WE), ethanol extract (EE), residue water extract (RWE) and petroleum ether (PF), ethyl acetate (EF), n-BuOH (BF) and water (WF) fractions of the ethanol extract from Eupatorium Lindley DC were investigated for the first time. Total phenolics content, DPPH radical scavenging activities, superoxide radical scavenging activities, total reduction capability, and ferrous ions chelating activities were determined for all the extracts and fractions. The results showed that all the extracts and fractions exhibited antioxidant activities with different magnitudes of potency. Among all the samples, WE and RWE exhibited the best antioxidant capacities, the BF also exhibited high antioxidant abilities in all tests except for the metal chelating activity, while the other extracts and fractions were relatively weak antioxidants. The BF had the highest total phenolics contents in all extracts and fractions, and the WE and RWE were found to be rich in tannins. Furthermore, the content of total phenolics showed good correlation with DPPH radical scavenging activity, superoxide anion radical scavenging activity, and the reducing power. Phenolic composition of all the extracts and fractions was identified and quantified by HPLC. The results indicate that the extracts of E. Lindley DC might be a useful potential source of natural antioxidant ingredients.
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Antioxidantes/química , Eupatorium/química , Fenóis/química , Extratos Vegetais/química , Quelantes/química , Sequestradores de Radicais Livres/químicaRESUMO
Vina-ginsenoside R4 (VGN4) is the first example of protopanaxatriol saponin possessing sugar chains located at C-3 and C-20 of aglycone. However, to the best of our knowledge, no report has been published on the neuroprotective effect of VGN4. In the present work, we investigated the neuroprotective effect of VGN4 against 6-hydroxydopamine (6-OHDA)-induced toxicity and its potential mechanism. Pretreatment of PC12 cells with VGN4 attenuated 6-OHDA-induced cell damage and cell apoptosis, which was correlated with the decrease of reactive oxygen species and the increase of antioxidant enzyme activities including superoxide dismutase and catalase. In addition, VGN4 markedly decreased nuclear translation of the nuclear factor-κB and PI3K/Akt/GSK/3ß signaling pathway including p85, PDK1, Akt, and GSK-3ß. Further studies revealed that PI3K siRNA attenuated the neuroprotective effect of VGN4 on caspase-3 activity. These data indicate that VGN4 might have the potential to be developed as a new neuroprotective functional food.
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Medicamentos de Ervas Chinesas/farmacologia , Ginsenosídeos/farmacologia , Glicogênio Sintase Quinase 3 beta/metabolismo , Síndromes Neurotóxicas/metabolismo , Panax/química , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Animais , Glicogênio Sintase Quinase 3 beta/genética , Humanos , Fármacos Neuroprotetores/farmacologia , Síndromes Neurotóxicas/tratamento farmacológico , Síndromes Neurotóxicas/etiologia , Síndromes Neurotóxicas/genética , Oxidopamina/toxicidade , Células PC12 , Fosfatidilinositol 3-Quinases/genética , Folhas de Planta/química , Proteínas Proto-Oncogênicas c-akt/genética , Ratos , Transdução de Sinais/efeitos dos fármacosRESUMO
Six new pentacyclic triterpenoid saponins, centelloside F (1), centelloside G (2), 11-oxo-asiaticoside B (3), 11-oxo-madecassoside (4), 11(ß)-methoxy asiaticoside B (5), and 11(ß)-methoxy madecassoside (6), along with seven known ones, asiaticoside (7), asiaticoside B (8), madecassoside (9), centellasaponin A (10), isoasiaticoside (11), scheffoleoside A (12), and centelloside E (13), were separated from the 80% MeOH extract of the whole plant of Centella asiatica, which has been used as a medicinal plant and is now commercially available as a diatery supplement in many countries. Compounds 1 and 2, 3 and 4, and 5 and 6 are three pairs of isomers with oleanane- or ursane-type triterpenes as aglycones. The chemical structures of the new triterpene saponins were fully characterized by extensive analysis of their nuclear magnetic resonance and high-resolution electrospray ionization mass spectrometry data. The protective effects of compounds 1-13 on PC12 cells induced by 6-OHDA were screened, and compound 3 displayed the best neuroprotective effect, with 91.75% cell viability at the concentration of 100 µM. Moreover, compound 3 also attenuated cell apoptosis and increased the mRNA expression of antioxidant enzymes, including superoxide dismutase and catalase. Additionally, compound 3 activated the phosphatidylinositol 3-kinase/Akt pathway, including PDK1, Akt, and GSK-3ß. These findings suggested that triterpene saponins from C. asiatica were worthy of further biological research to develop new neuroprotective agents.
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Centella/química , Fármacos Neuroprotetores/farmacologia , Ácido Oleanólico/análogos & derivados , Extratos Vegetais/química , Triterpenos/farmacologia , Animais , Apoptose/efeitos dos fármacos , Catalase/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Glicogênio Sintase Quinase 3 beta/metabolismo , Neurônios/citologia , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Fármacos Neuroprotetores/química , Ácido Oleanólico/química , Ácido Oleanólico/farmacologia , Células PC12 , Extratos Vegetais/farmacologia , Ratos , Superóxido Dismutase/metabolismo , Triterpenos/químicaRESUMO
BACKGROUND: The leaves and roots of Panax ginseng are rich in ginsenosides. However, the chemical compositions of the leaves and roots of P. ginseng differ, resulting in different medicinal functions. In recent years, the aerial parts of members of the Panax genus have received great attention from natural product chemists as producers of bioactive ginsenosides. The aim of this study was the isolation and structural elucidation of novel, minor ginsenosides in the leaves of P. ginseng and evaluation of their antiinflammatory activity in vitro. METHODS: Various chromatographic techniques were applied to obtain pure individual compounds, and their structures were determined by nuclear magnetic resonance and high-resolution mass spectrometry, as well as chemical methods. The antiinflammatory effect of the new compounds was evaluated on lipopolysaccharide-stimulated RAW 264.7 cells. RESULTS AND CONCLUSIONS: Two novel, minor triterpenoid saponins, ginsenoside LS1 (1) and 5,6-didehydroginsenoside Rg3 (2), were isolated from the leaves of P. ginseng. The isolated compounds 1 and 2 were assayed for their inhibitory effect on nitric oxide production in LPS-stimulated RAW 264.7 cells, and Compound 2 showed a significant inhibitory effect with IC50 of 37.38 µM compared with that of NG-monomethyl-L-arginine (IC50 = 90.76 µM). Moreover, Compound 2 significantly decreased secretion of cytokines such as prostaglandin E2 and tumor necrosis factor-α. In addition, Compound 2 significantly suppressed protein expression of inducible nitric oxide synthase and cyclooxygenase-2. These results suggested that Compound 2 could be used as a valuable candidate for medicinal use or functional food, and the mechanism is warranted for further exploration.
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Wheat bran-derived polysaccharides have attracted particular attention due to their immunomodulatory effects. However, the molecular mechanisms underlying their functions are poorly understood. The current study was designed to examine the effect of wheat bran polysaccharide (WBP) on RAW 264.7 cells and the underlying signaling pathways, which have not been explored. In addition, we also investigated the immuno-enhancement effects of WBP on cyclophosphamide (CTX)-induced immunosuppression in mice. WBP significantly increased the concentrations of intracellular nitric oxide (NO) and cytokines such as prostaglandin E2 (PGE2) and tumor necrosis factor-α (TNF-α) in RAW 264.7 cells. The result of RT-PCR analysis indicated that WBP also enhanced inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and TNF-α expression. Further analyses demonstrated that WBP rapidly activated phosphorylated p38 mitogen-activated protein kinase (MAPK) and the transcriptional activities of activator protein-1 (AP-1) and nuclear factor (NF)-κB via toll-like receptor 4 (TLR4). Furthermore, in vivo experiments revealed that WBP increased the spleen and thymus indices significantly, and markedly promoted the production of the serum cytokines IL-2 and IFN-γ in CTX-induced immunosuppressed mice. Taken together, these results suggest that WBP can improve immunity by enhancing immune function, and could be explored as a potential immunomodulatory agent in functional food.
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ETHNOPHARMACOLOGICAL RELEVANCE: Mahonia bealei has a long history of medical use in traditional Chinese medicine for the treatment of inflammatory-associated diseases. Despite numerous phytochemical and pharmacological studies, there is a lack of systematic studies to understand the cellular and molecular mechanisms of the anti-inflammatory activity of this plant. AIM OF STUDY: This study aimed to evaluate the anti-inflammatory activity of the dichloromethane fraction from M. bealei leaves (MBL-CH). MATERIALS AND METHODS: RAW 264.7 cells were pretreated with different concentrations of MBL-CH for 30min prior to treatment with 1µg/ml of lipopolysaccharide (LPS). The nuclear factor κB (NF-κB) pathway and subsequent production of inflammatory mediators, such as nitric oxide (NO), prostaglandin E2 (PGE2), and tumour necrosis factor (TNF)-α were investigated. Furthermore, the in vivo mouse model of LPS-induced acute lung injury (ALI) was employed to study the anti-inflammatory effects of MBL-CH. RESULTS: Pre-treatment with MBL-CH significantly inhibited the LPS-stimulated secretion of NO, PGE2, and TNF-α into the culture medium, as well as the mRNA levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and TNF-α, which were associated with a reduction in the phosphorylation of IκBα, Akt, and PI3K and inhibition of the transcriptional activity of NF-κB. Furthermore, in vivo experiments revealed that MBL-CH attenuated LPS-stimulated lung inflammation in mice. CONCLUSION: Taken together, our findings indicate that MBL-CH attenuates LPS-stimulated inflammatory responses in macrophages by blocking NF-κB activation through interference with activation of the PI3K/Akt pathway, providing scientific evidence that the plant can be employed in traditional remedies.
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Anti-Inflamatórios/farmacologia , Pulmão/efeitos dos fármacos , Macrófagos Peritoneais/efeitos dos fármacos , Mahonia/química , Cloreto de Metileno/química , Extratos Vegetais/farmacologia , Folhas de Planta/química , Pneumonia/prevenção & controle , Solventes/química , Animais , Anti-Inflamatórios/isolamento & purificação , Citocinas/metabolismo , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Mediadores da Inflamação/metabolismo , Lipopolissacarídeos/farmacologia , Pulmão/imunologia , Pulmão/metabolismo , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos Peritoneais/imunologia , Macrófagos Peritoneais/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , NF-kappa B/metabolismo , Fosfatidilinositol 3-Quinase/metabolismo , Fitoterapia , Extratos Vegetais/isolamento & purificação , Plantas Medicinais , Pneumonia/induzido quimicamente , Pneumonia/imunologia , Pneumonia/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Células RAW 264.7 , Transdução de Sinais/efeitos dos fármacos , Fatores de TempoRESUMO
Apigenin-7-O-ß-D-glucuronide (AG), an active flavonoid derivative isolated from the agricultural residue of Juglans sigillata fruit husks, possesses multiple pharmacological activities, including anti-oxidant, anti-complement, and aldose reductase inhibitory activities. To date, no report has identified the anti-inflammatory mechanisms of AG. This study was therefore designed to characterize the molecular mechanisms of AG on lipopolysaccharide (LPS)-induced inflammatory cytokines in RAW 264.7 cells and on endotoxin-induced shock in mice. AG suppressed the release of nitric oxide (NO), prostaglandin E2 (PGE2), and tumour necrosis factor-α (TNF-α) in LPS-stimulated RAW 264.7 macrophages in a dose-dependent manner without affecting cell viability. Additionally, AG suppressed LPS-induced mRNA expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and TNF-α. AG treatment decreased the translocation of c-Jun into the nucleus, and decreased activator protein-1 (AP-1)-mediated luciferase activity through the inhibition of both p38 mitogen-activated protein kinase (MAPK) and extracellular signal-regulated kinase (ERK) phosphorylation. Consistent with the in vitro observations, AG protected mice from LPS-induced endotoxin shock by inhibiting proinflammatory cytokine production. Taken together, these results suggest that AG may be used as a source of anti-inflammatory agents as well as a dietary complement for health promotion.