RESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Baoyuan Decoction (BYD) was initially recorded in the classic of "Bo Ai Xin Jian" in the Ming dynasty. It is traditionally used for treating weakness and cowardice, and deficiency of vital energy. In researches related to anti-fatigue effects, the reciprocal regulation of AMPK and circadian clocks likely plays an important role in anti-fatigue mechanism, while it has not yet been revealed. Therefore, we elucidated the anti-fatigue mechanism of BYD through AMPK/CRY2/PER1 pathway. AIM OF THE STUDY: To investigate the effect and mechanism of BYD in reducing fatigue, using pharmacodynamics, network pharmacology and transcriptomics through the AMPK/CRY2/PER1 signaling pathway. MATERIALS AND METHODS: Firstly, the chemical constituents of BYD were qualitatively identified by UHPLC-Q-Exactive Orbitrap/MS, establishing a comprehensive strategy with an in-house library, Xcalibur software and Pubchem combined. Secondly, a Na2SO3-induced fatigue model and 2,2'-Azobis (2-methylpropionamidine) dihydrochloride (AAPH)-induced oxidative stress model were developed to evaluate the anti-fatigue and anti-oxidant activities of BYD using AB zebrafish. The anti-inflammatory activity of BYD was evaluated using CuSO4-induced and tail cutting-induced Tg (lyz: dsRed) transgenic zebrafish inflammation models. Then, target screening was performed by Swiss ADME, GeneCards, OMIM and DrugBank databases, the network was constructed using Cytoscape 3.9.0. Transcriptome and network pharmacology technology were used to investigate the related signaling pathways and potential mechanisms after treatment with BYD, which were verified by real-time quantitative PCR (RT-qPCR). RESULTS: In total, 114 compounds from the water extract of BYD were identified as major compounds. Na2SO3-induced fatigue model and AAPH-induced oxidative stress model indicated that BYD has significant anti-fatigue and antioxidant effects. Meanwhile, BYD showed significant anti-inflammatory effects on CuSO4-induced and tail cutting-induced zebrafish inflammation models. The KEGG result of network pharmacology showed that the anti-fatigue function of BYD was mainly effected through AMPK signaling pathway. Besides, transcriptome analysis indicated that the circadian rhythm, AMPK and IL-17 signaling pathways were recommended as the main pathways related to the anti-fatigue effect of BYD. The RT-qPCR results showed that compared with a model control group, the treatment of BYD significantly elevated the expression mRNA of AMPK, CRY2 and PER1. CONCLUSION: Herein, we identified 114 chemical constituents of BYD, performed zebrafish activity validation, while demonstrated that BYD can relieve fatigue by AMPK/CRY2/PER1 signaling pathway through network pharmacology and transcriptome.
Assuntos
Proteínas Quinases Ativadas por AMP , Amidinas , Medicamentos de Ervas Chinesas , Animais , Peixe-Zebra , Estresse Oxidativo , Fadiga/tratamento farmacológico , Medicamentos de Ervas Chinesas/farmacologia , Medicamentos de Ervas Chinesas/uso terapêutico , Inflamação/tratamento farmacológico , Antioxidantes , Transdução de Sinais , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêuticoRESUMO
Five kinds of exopolysaccharides (EPS) were obtained by fermentation of Scleroderma areolatum Ehrenb. with sucrose, glucose, maltose, lactose, and fructose as carbon sources. Antioxidant abilities of the obtained EPSs were evaluated by inhibiting AAPH, HO·, and glutathione (GS·) induced oxidation of DNA and quenching 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonate) cationic radical (ABTS· and galvinoxyl radicals. The effects of carbon sources on the antioxidant properties of EPSs could be examined. The results showed that five EPSs can effectively inhibit radicals induced oxidation of DNA, and the thiobarbituric acid reactive substances (TBARS) percentages were 44.7%-80.8%, 52.3%-77.5%, and 44.7%-73.3% in inhibiting AAPH, HO·, and GS· induced oxidation of DNA, respectively. All five EPSs could scavenge ABTS· and galvinoxyh, and exhibit superior activity in scavenging free radicals. Antioxidant abilities of EPS with fructose as carbon source were highest among five EPS.
Assuntos
Amidinas , Antioxidantes , Basidiomycota , Benzotiazóis , Carbono , Ácidos Sulfônicos , Antioxidantes/farmacologia , Antioxidantes/química , DNA/química , Frutose , Sequestradores de Radicais Livres/farmacologiaRESUMO
BACKGROUND: Butea superba Roxb. (B. superba), is an herbal plant traditionally used for rejuvenation. Additionally, there have been reports on its antioxidant properties. Low-density lipoproteins (LDL) oxidation is the leading cause of cardiovascular diseases (CVDs). Natural products with antioxidant properties have the potential to inhibit LDL oxidation. However, no work has been done about the anti-isolated human LDL oxidation of B. superba extract (BSE). This study aimed to investigate the antioxidant potential of BSE and its ability to prevent isolated human (LDL) oxidation induced by free radical agents. METHODS: The antioxidant properties were investigated by antioxidant assays, including 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azinobis-(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS), ferric reducing ability power (FRAP), nitric oxide (NO) and peroxynitrite scavenging assay. More so, anti-isolated human LDL oxidation activities were evaluated by 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH) and 3-morpholinosydnonimine hydrochloride (SIN-1) induced LDL oxidation assay. RESULTS: BSE exhibited a significant (p < 0.05) antioxidant activity in all the test systems, demonstrating its potential as a potent free radical scavenger. It displayed scavenging effects on DPPH (p < 0.05; IC50 = 487.67 ± 21.94 µg/ml), ABTS (p < 0.05; IC50 = 30.83 ± 1.29 µg/ml). Furthermore, it generated significantly (p < 0.05) increased antioxidant capacity in a dose-dependent manner in FRAP assay and exhibited significantly (p < 0.01) higher percent NO scavenging activity than gallic acid. Besides, BSE at 62.5 µg/ml exhibited a considerable percent peroxynitrite scavenging of 71.40 ± 6.59% after a 2 h period. Moreover, BSE demonstrated anti-isolated human LDL oxidation activity induced by AAPH and SIN-1 (p < 0.05) and revealed scavenging activity similar to ascorbic acid (p > 0.05). Identifying the main constituents of BSE revealed the presence of genistein, daidzein, and biochanin A through Liquid Chromatography-Mass Spectrometer/Mass Spectrometer (LC-MS/MS) analysis. CONCLUSION: This is the first report that the presence of isoflavones in BSE could play an important role in its antioxidation and isolated human LDL oxidation scavenging properties. These findings suggest the potential for developing antioxidant herbal supplements. However, further studies must be investigated, including efficacious and safe human dosages.
Assuntos
Amidinas , Antioxidantes , Benzotiazóis , Butea , Ácidos Sulfônicos , Humanos , Antioxidantes/farmacologia , Antioxidantes/química , Butea/química , Cromatografia Líquida , Ácido Peroxinitroso , Extratos Vegetais/farmacologia , Extratos Vegetais/química , Espectrometria de Massas em Tandem , Óxido Nítrico , Radicais LivresRESUMO
Candida tropicalis is an emerging pathogen with a high mortality rate due to its virulence factors, including biofilm formation, that has important repercussions on the public health system. The ability of C. tropicalis to form biofilms, which are potentially more resistant to antifungal drugs and the consequent increasing antimicrobial resistance, highlights an urgent need for the development of novel antifungal. The present study analyzed the antibiofilm capacity of the arylamidine T-2307 on two strains of Candida tropicalis. Antimicrobial activity and time-killing assays were performed to evaluate the anticandidal effects of T-2307, the antibiofilm ability on biomass inhibition and eradication was evaluated by the crystal violet (CV) method. Furthermore, in Galleria mellonella infected larvae an increased survival after pre-and post- treatment with T-2307 was observed. The MTT test was used to determine the viability of immortalized human prostate epithelial cells (PNT1A) after exposure to different concentrations of T-2307. Levels of interleukin IL-4, IL-8, IL-10 were quantified after Candida infection of PNT1A cells and treatment. Active doses of T-2307 did not affect the viability of PNT1A cells, and drug concentrations of 0.005 or 0.01 µg mL-1 inhibited the secretion of inflammatory cytokines. Taken together, these results provide new information on T-2307, indicating this drug as a new and promising alternative therapeutic option for the treatment of Candida infections.
Assuntos
Antifúngicos , Candidíase , Masculino , Animais , Humanos , Antifúngicos/farmacologia , Candida tropicalis/fisiologia , Amidinas/farmacologia , Candidíase/tratamento farmacológico , Candidíase/microbiologia , Biofilmes , Testes de Sensibilidade MicrobianaRESUMO
Redox imbalance can lead to irreversible damages to biological functions. In this context, rutin stands out for its antioxidant potential. The objective of this study was to evaluate the acute and chronic effect of rutin on the hepatic redox imbalance. The study was performed according to three different protocols. First, healthy male Swiss mice were divided into two groups: control and rutin, the second of which received chronic oral supplementation of rutin (10 mg/kg). The second involved evaluation of the generation of reactive oxygen species (ROS) by HepG2 cells, incubated or not with rutin (20 and 40 µg/mL) for 3 h. The final protocol involved assessment of the acute effect of rutin (10 mg/kg) in mice with oxidative stress induced by 2,2'-azobis(2-methylpropionamidine) dihydrochloride (ABAP). After the in vivo treatments, the livers were collected to analyze the oxidative damage by thiol, and the antioxidant defense by catalase, superoxide dismutase, and glutathione peroxidase. In the HepG2 cells, the following probes were employed to assess the ROS production: dichlorofluorescein, MitoSOX, dihydroethidium, and Amplex Red. Rutin administered chronically improved the antioxidant defense in healthy animals, and when administered acutely both inhibited the increased production of ROS in HepG2 cells and improved the redox imbalance parameters in mice with induced oxidative stress. This study suggests rutin as a protective agent for restoration of hepatic redox homeostasis in redox injury induced by ABAP in Swiss mice and HelpG2 cells.
Assuntos
Antioxidantes , Rutina , Amidinas , Animais , Antioxidantes/metabolismo , Antioxidantes/farmacologia , Células Hep G2 , Humanos , Fígado/metabolismo , Masculino , Camundongos , Oxirredução , Estresse Oxidativo , Espécies Reativas de Oxigênio/metabolismo , Rutina/metabolismo , Rutina/farmacologiaRESUMO
In this project, the new catalyst copper defines as Fe3O4@Pectin@(CH2)3-Acetamide-Cu(II) was successfully manufactured and fully characterized by different techniques, including FT-IR, XRD, TEM, FESEM, EDX, VSM, TGA, and ICP analysis. All results showed that copper was successfully supported on the polymer-coated magnetic nanoparticles. One of the most important properties of a catalyst is the ability to be prepared from simple materials such as pectin that's a biopolymer that is widely found in nature. The catalytic activity of Fe3O4@Pectin@(CH2)3-Acetamide-Cu(II) was examined in a classical, one pot, and the three-component reaction of terminal alkynes, alkyl halides, and sodium azide in water and observed, proceeding smoothly and completed in good yields and high regioselectivity. The critical potential interests of the present method include high yields, recyclability of catalyst, easy workup, using an eco-friendly solvent, and the ability to sustain a variety of functional groups, which give economical as well as ecological rewards. The capability of the nanocomposite was compared with previous works, and the nanocomposite was found more efficient, economical, and reproducible. Also, the catalyst can be easily removed from the reaction solution using an external magnet and reused for five runs without reduction in catalyst activity.
Assuntos
Nanopartículas de Magnetita , Bases de Schiff , Amidinas , Química Click , Cobre/química , Nanopartículas de Magnetita/química , Pectinas/química , Espectroscopia de Infravermelho com Transformada de Fourier , Triazóis/químicaRESUMO
A series of 6-amidinobenzothiazoles, linked via phenoxymethylene or directly to the 1,2,3-triazole ring with a p-substituted phenyl or benzyl moiety, were synthesised and evaluated in vitro against four human tumour cell lines and the protozoan parasite Trypanosoma brucei. The influence of the type of amidino substituent and phenoxymethylene linker on antiproliferative and antitrypanosomal activities was observed, showing that the imidazoline moiety had a major impact on both activities. Benzothiazole imidazoline 14a, which was directly connected to N-1-phenyl-1,2,3-triazole, had the most potent growth-inhibitory effect (IC50 = 0.25 µM) on colorectal adenocarcinoma (SW620), while benzothiazole imidazoline 11b, containing a phenoxymethylene linker, exhibited the best antitrypanosomal potency (IC90 = 0.12 µM). DNA binding assays showed a non-covalent interaction of 6-amidinobenzothiazole ligands, indicating both minor groove binding and intercalation modes of DNA interaction. Our findings encourage further development of novel structurally related 6-amidino-2-arylbenzothiazoles to obtain more selective anticancer and anti-HAT agents.
Assuntos
Antiprotozoários/síntese química , Benzotiazóis/síntese química , Substâncias Intercalantes/síntese química , Trypanosoma brucei brucei/efeitos dos fármacos , Amidinas/química , Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Antiprotozoários/farmacologia , Benzotiazóis/farmacologia , Proliferação de Células/efeitos dos fármacos , Neoplasias Colorretais/tratamento farmacológico , DNA/química , Avaliação Pré-Clínica de Medicamentos , Humanos , Imidazolinas/química , Substâncias Intercalantes/farmacologia , Conformação de Ácido Nucleico , Relação Estrutura-Atividade , Triazóis/químicaRESUMO
A novel polysaccharide (MFP1P) was isolated from Fructus Mori, followed by purification via DEAE-52 cellulose and 27 % ethanol fraction. The MFP1P had the molecular weight of 56.78 kDa and the total sugar content of 93.32±0.54 %. And the MFP1P is mainly composed of glucose, galactose, galacturonic acid and mannose with molar ratio of 66.62 %, 13.94 %, 18.24 % and 1.20 %, respectively. MFP1P was mainly composed of â3)-α-D-Gal (1â, ß-D-Man-(1â and â6)-α-D-Glc (1â glycosidic bond and showed a spherical chain conformation with uniform distribution in solution. The MFP1P exhibited great antioxidant activity with oxygen-free radical absorption capacity (ORAC) values of 291.63±6.81 µmol TE/g and MDA IC50 of 0.289±0.022 mg/mL.
Assuntos
Antioxidantes/química , Frutas/química , Fígado/efeitos dos fármacos , Morus/química , Oxidantes/antagonistas & inibidores , Polissacarídeos/química , Amidinas/antagonistas & inibidores , Amidinas/química , Animais , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Sequência de Carboidratos , Fracionamento Químico/métodos , Misturas Complexas/química , Galactose/química , Galactose/isolamento & purificação , Glucose/química , Glucose/isolamento & purificação , Ácidos Hexurônicos/química , Ácidos Hexurônicos/isolamento & purificação , Fígado/metabolismo , Malondialdeído/antagonistas & inibidores , Malondialdeído/metabolismo , Manose/química , Manose/isolamento & purificação , Camundongos , Peso Molecular , Oxidantes/química , Extratos Vegetais/química , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologiaRESUMO
Tea is one of the most popular beverages in the world. Camellia sinensis tea (CST) or green tea is widely regarded as a potent antioxidant. In Thailand, Pluchea indica (L.) Less. tea (PIT) has been commercially available as a health-promoting drink. This study focused on free radical scavenging activities of PIT, and its ability to protect isolated human low-density lipoproteins (LDL) from oxidation by chemical agents. A preliminary study to investigate the antioxidant nature of PIT was undertaken. These included common antioxidant assays involving 2,2-Diphenyl-1-picrylhydrazyl (DPPH), 2,2-azinobis-(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS), hypochlorous acid (HOCl), and its potential to scavenge peroxynitrite. In separated experiments, isolated human LDL was challenged with either 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH), copper (Cu2+), or 3-Morpholinosydnonimine hydrochloride (SIN-1) to induce LDL oxidation. PIT exhibited antioxidant activity in all test systems and performed significantly better than CST in both DPPH (P < 0.05; IC50PIT = 245.85 ± 15.83 and CST = 315.41 ± 24.18 µg/ml) and peroxynitrite scavenging assays. PIT at 75 µg/ml almost fully prevented the peroxynitrite over a 5 h period. Moreover, it displayed similar properties to CST during the antioxidation of isolated human LDL using AAPH, Cu2+, SIN-1, and hypochlorous acid scavenging assays. However, it revealed a significantly lower ABTS scavenging activity than CST (P < 0.05; IC50PIT = 30.47 ± 2.20 and CST = 21.59 ± 0.67 µg/ml). The main constituents of the PIT were identified using LC-MS/MS. It contained 4-O-caffeoylquinic acid (4-CQ), 5-O-caffeoylquinic acid (5-CQ), 3,4-O-dicaffeoylquinic acid (3,4-CQ), 3,5-O-dicaffeoylquinic acid (3,5-CQ), and 4,5-O-dicaffeoylquinic acid (4,5-CQ). In conclusion, caffeoyl derivatives in PIT could play an important role in potent antioxidant properties. So, it may be further developed to be antioxidant beverages for preventing atherosclerosis and cardiovascular diseases associated with oxidative stress.
Assuntos
Asteraceae/química , Camellia sinensis/química , Sequestradores de Radicais Livres/farmacologia , Lipoproteínas LDL/metabolismo , Amidinas/farmacologia , Benzotiazóis/química , Compostos de Bifenilo/química , Cobre/farmacologia , Humanos , Ácido Hipocloroso/química , Molsidomina/análogos & derivados , Molsidomina/farmacologia , Óxido Nítrico/metabolismo , Oxirredução , Ácido Peroxinitroso/metabolismo , Picratos/química , Ácidos Sulfônicos/químicaRESUMO
Fucoidan is a fucose-enriched polysaccharide, obtained from brown algae, with demonstrated antioxidant properties. However, traditional extraction methods using water or chemical-based extraction methods have reduced yield and produced hazardous by-products. In this study, we isolated fucoidan at a high yield using enzyme-assisted extraction; the Celluclast enzyme assisted extract of Undaria pinnatifida sporophylls (FCUS). To examine the antioxidant properties of FCUS, oxidative stress was induced with 2,2'-azobis (2-methylpropionamidine) dihydrochloride (AAPH) in Vero cells and zebrafish model. FCUS was composed of 30.4% sulfate and 52.3% fucose. Pre-treatment of Vero cells with FCUS dose dependently inhibited AAPH-induced reactive oxygen species (ROS) production. Moreover, FCUS remarkably reduced cell death, ROS generation, and lipid peroxidation production in zebrafish larvae. Overall, these findings indicate that the sulfate-rich fucoidan of FCUS, obtained with an eco-friendly process, could be implemented as a beneficial antioxidant agent in the functional food industry.
Assuntos
Antioxidantes/química , Estresse Oxidativo/efeitos dos fármacos , Polissacarídeos/química , Undaria/química , Amidinas/toxicidade , Animais , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Chlorocebus aethiops , Peroxidação de Lipídeos/efeitos dos fármacos , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologia , Espécies Reativas de Oxigênio/química , Células Vero , Peixe-ZebraRESUMO
A novel and simple method was established for the extraction and determination of seven compounds in Anemarrhena asphodeloides Bge. using silica gel-based vortex-homogenized matrix solid-phase dispersion and ultra-high performance liquid chromatography quadrupole-time of-flight mass spectrometer. The conditions for the extraction were optimized. Silica gel was used as the dispersant, 50% methanol-water was selected as an elution solvent and the grinding time was 3 min. Compared with the traditional ultrasonic-assisted extraction, the developed method was rapid and efficient. In order to screen potential antioxidants, extract dealing with the optimized method was applied to a polyamide chromatography column and a D-101 macroporous resin column. Fr.2.2 showed the highest antioxidant activities with the most content of flavonoid. A total of 25 peaks were identified from the active fraction. A 2,2'-diphenyl-1-picrylhydrazyl ultra-high performance liquid chromatography coupled with mass spectrometry approach was adopted for the rapid and exact screening and identification of antioxidant compounds. It indicated that flavonoids exhibited potential antioxidant activities. The antioxidant activities of nine monomeric compounds in vivo were tested. Structure-activity relationships were discussed. Five flavonoids with the concentration of 500 µg/mL would reduce the oxidative stress of PC12 cells that were induced with 2,2'-azobis[2-methylpropionamidine] dihydrochloride.
Assuntos
Anemarrhena/química , Antioxidantes/análise , Flavonoides/análise , Extratos Vegetais/isolamento & purificação , Extração em Fase Sólida , Amidinas/antagonistas & inibidores , Amidinas/farmacologia , Animais , Antioxidantes/farmacologia , Relação Dose-Resposta a Droga , Flavonoides/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Células PC12 , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Ratos , Sílica Gel/químicaRESUMO
PURPOSE: Laparoscopic inguinal hernia repair (IHR) may lead to early postoperative pain. Therefore, opioid and non-opioid analgesic agents are often administered in the post-anesthesia care unit (PACU). To reduce the postoperative cumulative need of analgesic medication, as well as to accelerate the physical recovery time, the transversus abdominis plane (TAP) block has recently been studied. The TAP block is a regional anesthesia technique. Even though there is evidence about the efficacy of the block used in procedure such as an open inguinal hernia repair, the evidence regarding its use for the TAPP (transabdominal preperitoneal) technique remains low. We aim to provide more sufficient evidence regarding this topic. METHODS: A monocentric retrospective observational study investigating the effect of the TAP block prior to primary IHR in TAPP technique was conducted. The data of 838 patients who were operated on using this technique from June 2007 to February 2019 were observed. 72 patients were excluded because of insufficient information regarding their analgesic medication protocol. The patients' data were taken from their files. RESULTS: The patients in the TAP block group (n = 364) did not differ statistically significantly compared to the control group (n = 402) in terms of gender, BMI and age. Individuals of the TAP block group experienced less postoperative pain in the PACU (p < 0.001) and received less analgesic medication (morphine, oxycodone, piritramide, acetaminophen; p < 0.001). CONCLUSION: We assume that the TAP block is a sufficient approach to reduce postoperative pain and analgesic medication administration for IHR in TAPP technique.
Assuntos
Músculos Abdominais/efeitos dos fármacos , Amidinas/uso terapêutico , Anestesia Local/métodos , Hérnia Inguinal/tratamento farmacológico , Bloqueio Nervoso/métodos , Músculos Abdominais/cirurgia , Amidinas/farmacologia , Feminino , Hérnia Inguinal/cirurgia , Humanos , Masculino , Estudos RetrospectivosRESUMO
Phenolamines and flavonoids are two important components in bee pollen. There are many reports on the bioactivity of flavonoids in bee pollen, but few on phenolamines. This study aims to separate and characterize the flavonoids and phenolamines from rape bee pollen, and compare their antioxidant activities and protective effects against oxidative stress. The rape bee pollen was separated to obtain 35% and 50% fractions, which were characterized by HPLC-ESI-QTOF-MS/MS. The results showed that the compounds in 35% fraction were quercetin and kaempferol glycosides, while the compounds in 50% fraction were phenolamines, including di-p-coumaroyl spermidine, p-coumaroyl caffeoyl hydroxyferuloyl spermine, di-p-coumaroyl hydroxyferuloyl spermine, and tri-p-coumaroyl spermidine. The antioxidant activities of phenolamines and flavonoids were evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS), and ferric reducing antioxidant power (FRAP) assays. It was found that the antioxidant activity of phenolamines was significantly higher than that of flavonoids. Moreover, phenolamines showed better protective effects than flavonoids on HepG2 cells injured by AAPH. Furthermore, phenolamines could significantly reduce the reactive oxygen species (ROS), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, and increase the superoxide dismutase (SOD) and glutathione (GSH) levels. This study lays a foundation for the further understanding of phenolamines in rape bee pollen.
Assuntos
Antioxidantes/química , Glicosídeos/química , Quempferóis/química , Pólen/química , Quercetina/química , Espermidina/química , Espermina/química , Alanina Transaminase/genética , Alanina Transaminase/metabolismo , Amidinas/antagonistas & inibidores , Amidinas/farmacologia , Animais , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Aspartato Aminotransferases/genética , Aspartato Aminotransferases/metabolismo , Abelhas , Benzotiazóis/antagonistas & inibidores , Benzotiazóis/química , Compostos de Bifenilo/antagonistas & inibidores , Compostos de Bifenilo/química , Expressão Gênica/efeitos dos fármacos , Glutationa/genética , Glutationa/metabolismo , Glicosídeos/isolamento & purificação , Glicosídeos/farmacologia , Células Hep G2 , Humanos , Quempferóis/isolamento & purificação , Quempferóis/farmacologia , Oxidantes/antagonistas & inibidores , Oxidantes/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Picratos/antagonistas & inibidores , Picratos/química , Extratos Vegetais/química , Quercetina/isolamento & purificação , Quercetina/farmacologia , Espécies Reativas de Oxigênio/antagonistas & inibidores , Espécies Reativas de Oxigênio/química , Espermidina/análogos & derivados , Espermidina/isolamento & purificação , Espermidina/farmacologia , Espermina/análogos & derivados , Espermina/isolamento & purificação , Espermina/farmacologia , Ácidos Sulfônicos/antagonistas & inibidores , Ácidos Sulfônicos/química , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismoRESUMO
Erythrocytes exhibit high susceptibility to hemolysis in several pathologies due to the oxidation of cellular components. We hypothesized that annatto carotenoids improve the redox status of erythrocyte plasma membranes and promote a consequent increase in human erythrocyte resistance to hemolysis. The objective of this study was to evaluate whether food-grade annatto carotenoids can increase human erythrocyte resistance to hemolysis in vitro and ex vivo. For the in vitro experiment, erythrocytes from healthy volunteers were isolated and coincubated with bixin (BIX) or norbixin (NBIX) and 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH), glucose, or sodium nitrite (NaNO2) as hemolysis inducers. In the ex vivo study, healthy volunteers consumed a capsule containing BIX or NBIX (0.05â¯mg/kg body weight per day) or placebo for 7â¯days before blood sample collection. Their erythrocytes were isolated and incubated with AAPH, glucose, or NaNO2. In both the ex vivo and in vitro studies, erythrocytes were subjected to osmotic fragility tests. The activity of antioxidant enzymes, and reduced glutathione and lipid peroxidation levels in erythrocytes were also evaluated ex vivo. In vitro BIX and NBIX not only reduced erythrocyte membrane fragility induced by AAPH, glucose, or NaNO2 but also improved basal osmotic resistance in the micromole-per-liter range (Pâ¯<â¯.05). BIX and NBIX supplementation increased erythrocyte membrane resistance (Pâ¯<â¯.05), with BIX being more effective. Also, BIX and NBIX protected erythrocytes from lipid peroxidation and improved the cellular redox environment (Pâ¯<â¯.05). These results support the hypothesis that annatto carotenoids supplementation exerts antihemolytic properties by preventing the oxidative damage of human erythrocytes.
Assuntos
Antioxidantes/farmacologia , Carotenoides/farmacologia , Suplementos Nutricionais , Eritrócitos/efeitos dos fármacos , Corantes de Alimentos/farmacologia , Hemólise/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/farmacologia , Adolescente , Adulto , Amidinas , Antioxidantes/metabolismo , Bixaceae/química , Glucose , Glutationa/metabolismo , Voluntários Saudáveis , Humanos , Peroxidação de Lipídeos , Oxirredução , Nitrito de Sódio , Adulto JovemRESUMO
OBJECTIVES: T-2307, a novel arylamidine, shows broad-spectrum activity against pathogenic fungi, including Candida albicans. Ocular candidiasis is one of the serious complications associated with Candida bloodstream infection and is known to be refractory to conventional antifungal agents. The aim of the present study was to clarify the effectiveness of T-2307 against ocular candidiasis using a mouse model. METHODS: We evaluated ocular fungal burden in mice infected with C. albicans that received treatment with antifungal agents [T-2307, liposomal amphotericin B (LAMB) or fluconazole] for 3 consecutive days. We also assessed survival rates of mice after C. albicans infection followed by treatment for 7 consecutive days. In addition, ocular T-2307 concentrations and in vitro effectiveness against C. albicans biofilm formation were evaluated. RESULTS: The ocular fungal burdens were significantly reduced after T-2307 treatment compared with the control group (no treatment received) and were comparable with those observed following treatment with LAMB or fluconazole in both early- and late-phase treatment experiments. In addition, all of the mice treated with antifungal agents survived for 3 weeks after infection, whereas mice in the control group died within 3 days. The ocular T-2307 trough concentration was maintained above the MIC in the infected mice. An in vitro biofilm inhibition experiment showed that T-2307 suppressed C. albicans biofilm formation at the sub-MIC level, which was comparable with amphotericin B. CONCLUSIONS: Given these results in experimental disseminated candidiasis, T-2307 may be an effective treatment against the complication of ocular candidiasis.
Assuntos
Amidinas/uso terapêutico , Antifúngicos/uso terapêutico , Candida albicans/efeitos dos fármacos , Candidíase/complicações , Infecções Oculares/tratamento farmacológico , Infecções Oculares/microbiologia , Amidinas/farmacologia , Animais , Antifúngicos/farmacologia , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Candida albicans/patogenicidade , Candidíase/tratamento farmacológico , Contagem de Colônia Microbiana , Farmacorresistência Fúngica , Olho/microbiologia , Feminino , Rim/microbiologia , Camundongos Endogâmicos C57BL , Testes de Sensibilidade Microbiana , Organismos Livres de Patógenos EspecíficosRESUMO
The effects of selenium (Se) on the protein content, amino acid profile, secondary structure and subunit composition of soy proteins and its distribution were evaluated, as was the effect of peroxyl radicals produced by thermal decomposition of AAPH on the conformational changes of Se-enriched ß-conglycinin (S-7S). The Se biofortification ability of soy was very strong, 7S had strongest ability to incorporate Se, and lower amounts of inorganic Se existed in Se-enriched beans. Se could promote protein synthesis and thus improve the protein content, increase the total amino acid content with a decrease in cysteine, combine into low-molecular-weight proteins, and influence the secondary structure of soybean proteins. Se was involved in the relevant protein changes in surface hydrophobicity, intrinsic fluorescence, infrared absorption and solubility and played an antioxidant role as an effectual "protector" to reduce the influence of peroxyl radical oxidation on S-7S, thereby maintaining the structural rearrangement between aggregation and protein unfolding.
Assuntos
Amidinas/farmacologia , Antígenos de Plantas/química , Antígenos de Plantas/farmacologia , Globulinas/química , Globulinas/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Proteínas de Armazenamento de Sementes/química , Proteínas de Armazenamento de Sementes/farmacologia , Selênio/análise , Proteínas de Soja/química , Proteínas de Soja/farmacologia , Peso Molecular , Estrutura Secundária de ProteínaRESUMO
Oxidative stress affects all the structures of the human eye, particularly the retina and its retinal pigment epithelium (RPE). The RPE limits oxidative damage by several protective mechanisms, including the non-enzymatic antioxidant system zinc-metallothionein (Zn-MT). This work aimed to investigate the role of Zn-MT in the protection of RPE from the oxidative damage of reactive oxygen intermediates by analytical and biochemical-based techniques. The Zn-MT system was induced in an in vitro model of RPE cells and determined by elemental mass spectrometry with enriched isotopes and mathematical calculations. Induced-oxidative stress was quantified using fluorescent probes. We observed that 25, 50 or 100 µM of zinc induced Zn-MT synthesis (1.6-, 3.6- and 11.9-fold, respectively), while pre-treated cells with zinc (25, 50, and 100 µM) and subsequent 2,2'-Azobis(2-methylpropionamidine) dihydrochloride (AAPH) treatment increased Zn-MT levels in a lesser extent (0.8-, 2.1-, 6.1-fold, respectively), exerting a stoichiometric transition in the Zn-MT complex. Moreover, AAPH treatment decreased MT levels (0.4-fold), while the stoichiometry remained constant or slightly higher when compared to non-treated cells. Convincingly, induction of Zn-MT significantly attenuated oxidative stress produced by free radicals' generators. We conclude that the stoichiometry of Zn-MT plays an important role in oxidative stress response, related with cellular metal homeostasis.
Assuntos
Antioxidantes/farmacologia , Metalotioneína/fisiologia , Estresse Oxidativo/efeitos dos fármacos , Epitélio Pigmentado da Retina/metabolismo , Adulto , Amidinas/farmacologia , Linhagem Celular , Humanos , Peróxido de Hidrogênio/farmacologia , Metalotioneína/análise , Metalotioneína/biossíntese , Metalotioneína/química , Metalotioneína/metabolismo , Oxidantes/farmacologia , Oxirredução , Epitélio Pigmentado da Retina/efeitos dos fármacos , Zinco/farmacologiaRESUMO
The physicochemical characteristics and functional properties of pumpkin (Cucurbita maxima D. var. Cabello de Ángel) pectin obtained by cavitation facilitated extraction from pumpkin pulp have been evaluated and compared with commercial citrus and apple pectins. C. maxima pectin had an Mw value of 90 kDa and a high degree (72%) of esterification. The cytoprotective and antioxidant effects of citrus, apple and pumpkin pectin samples with different concentrations were studied in vitro in cell lines HT-29 (human colon adenocarcinoma) and MDCK1 (canine kidney epithelium). All pectin samples exhibited cytoprotective effect in HT-29 and MDCK1 cells after incubation with toxic concentrations of cadmium and mercury for 4 h. Pumpkin pectin increased the proliferation of cadmium-treated MDCK1 cells by 210%. The studied pectins also inhibited oxidative stress induced by 2,2'-azobis(2-methylpropionamidine) dihydrochloride (AAPH) in cell cultures, as determined by measuring the production of intracellular reactive species using dihydrochlorofluorescein diacetate (DCFH-DA). Pectin from pumpkin pomace had the highest (p < 0.05) protective effect against reactive oxygen species generation in MDCK1 cells induced by AAPH. Distinctive features of pumpkin pectin were highly branched RG-I regions, the presence of RG-II regions and the highest galacturonic acid content among the studied samples of pectins. This correlates with a considerable protective effect of C. maxima pectin against oxidative stress and cytotoxicity induced by heavy metal ions. Thus, C. maxima pectin can be considered as a source of new functional foods of agricultural origin.
Assuntos
Antioxidantes/farmacologia , Citrus/química , Cucurbita/química , Malus/química , Pectinas/farmacologia , Amidinas/toxicidade , Animais , Antioxidantes/química , Cádmio/toxicidade , Proliferação de Células/efeitos dos fármacos , Citoproteção , Cães , Células HT29 , Humanos , Células Madin Darby de Rim Canino , Mercúrio/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Pectinas/químicaRESUMO
Strawberry fruits are highly appreciated by consumers worldwide due to their bright red color, typical aroma, and juicy texture. While the biological activity of the complete fruit has been widely studied, the potential beneficial effects of the achenes (commonly named seeds) remain unknown. In addition, when raw fruit and achenes are consumed, the digestion process could alter the release and absorption of their phytochemical compounds, compromising their bioactivity. In the present work, we evaluated the protective effects against oxidative damage of nondigested and digested extracts from strawberry fruit and achenes in human hepatocellular carcinoma (HepG2) cells. For that purpose, cells were treated with different concentration of the extracts prior to incubation with the stressor agent, AAPH (2,2'-azobis(2-amidinopropane) dihydrochloride). Subsequently, intracellular accumulation of reactive oxygen species (ROS) and the percentage of live, dead, and apoptotic cells were determined. Our results demonstrated that all the evaluated fractions were able to counteract the AAPH-induced damage, suggesting that the achenes also present biological activity. The positive effects of both the raw fruit and achenes were maintained after the in vitro digestion process.
Assuntos
Antioxidantes/química , Antioxidantes/farmacologia , Fragaria/química , Hepatócitos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Amidinas/efeitos adversos , Antioxidantes/isolamento & purificação , Apoptose/efeitos dos fármacos , Frutas/química , Células Hep G2 , Hepatócitos/metabolismo , Humanos , Extratos Vegetais/isolamento & purificação , Espécies Reativas de Oxigênio/metabolismo , Sementes/químicaRESUMO
Given the limitations of current antileishmanial drugs and the utility of oral combination therapy for other infections, developing an oral combination against visceral leishmaniasis should be a high priority. In vitro combination studies with DB766 and antifungal azoles against intracellular Leishmania donovani showed that posaconazole and ketoconazole, but not fluconazole, enhanced DB766 potency. Pharmacokinetic analysis of DB766-azole combinations in uninfected Swiss Webster mice revealed that DB766 exposure was increased by higher posaconazole and ketoconazole doses, while DB766 decreased ketoconazole exposure. In L. donovani-infected BALB/c mice, DB766-posaconazole combinations given orally for 5 days were more effective than DB766 or posaconazole alone. For example, 81% ± 1% (means ± standard errors) inhibition of liver parasite burden was observed for 37.5 mg/kg of body weight DB766 plus 15 mg/kg posaconazole, while 37.5 mg/kg DB766 and 15 mg/kg posaconazole administered as monotherapy gave 40% ± 5% and 21% ± 3% inhibition, respectively. Combination index (CI) analysis indicated that synergy or moderate synergy was observed in six of nine combined dose groups, while the other three were nearly additive. Liver concentrations of DB766 and posaconazole increased in almost all combination groups compared to monotherapy groups, although many increases were not statistically significant. For DB766-ketoconazole combinations evaluated in this model, two were antagonistic, one displayed synergy, and one was nearly additive. These data indicate that the efficacy of DB766-posaconazole and DB766-ketoconazole combinations in vivo is influenced in part by the pharmacokinetics of the combination, and that the former combination deserves further consideration in developing new treatment strategies against visceral leishmaniasis.