RESUMO
Bisphenol A (BPA) is a well-known endocrine disruptor with several effects on mammalian systems and has been linked to diseases, such as cancer. Bisphenol S (BPS) emerged as a likely alternative to BPA in industrial production. Despite being well studied and exhibiting BPA-like toxic capacity, many effects are still being elucidated. The blood coagulation system is well controlled in an effort to minimize blood loss. To our knowledge, no study reported actions of bisphenols in this system. The aim of this work was to evaluate the effects of bisphenols on blood coagulation. Zebrafish were used to measure bleeding time. To assess possible mechanisms, platelet-rich plasma was incubated with both bisphenols in the presence of arachidonic acid. Prothrombin time (PT) and activated partial thromboplastin time (APTT) assays were performed in the presence of BPA and BPS. Alignment of human factor VII sequence was compared to zebrafish and docking simulations performed with FVIIa and bisphenols. An extended time was observed in BPA-treated but not BPS-treated animals in bleeding time; in PT, bisphenols showed no effect. APTT was increased in the highest concentration of bisphenols, with no effects in platelet aggregation, indicating interference with factor VII. Protein alignment showed that both proteins have well conserved residues, as those being required for interaction of FVIIa-BPA and FVIIa-BPS complexes, as shown in molecular docking. Taken together, these data show BPA and BPS as capable of interfering with the coagulation process via FVIIa.
Assuntos
Compostos Benzidrílicos , Peixe-Zebra , Animais , Compostos Benzidrílicos/toxicidade , Coagulação Sanguínea , Humanos , Simulação de Acoplamento Molecular , Fenóis/toxicidadeRESUMO
ABSTRACT: Marine algae are natural sources of macromolecules known as sulfated polysaccharides. This class of compounds has attracted the interest of Pharmaceutical Sciences due to its pharmacological anticoagulant, antiplatelet and antithrombotic properties. Therefore, this study evaluated the anticoagulant potential of sulfated polysaccharides extracted from the algae Penicillus capitatus. The extracted sulfated polysaccharides were purified, partially characterized and their anticoagulant activity was evaluated. The extraction process followed by ethanol precipitation resulted in five fractions. Among the analyzed fractions, F44 contained highest concentration of sulfated polysaccharides. After the purified fraction F23, F44 displayed in vitro anticoagulant activity in a time testing for activated partial thromboplastin time and prothrombin time. The preferential mechanism effect was based on interactions between thrombin and factor Xa. Additional studies on structure pharmacological are required to test the viability of the use of sulfated polysaccharides as therapeutic agents.
RESUMO: As algas marinhas são fontes naturais de macromoléculas conhecidas como polissacarídeos sulfatados. Esta classe de compostos atraiu o interesse das Ciências Farmacêuticas devido às suas propriedades farmacológicas como anticoagulante, antiplaquetária e antitrombótica. Portanto, este estudo tem como objetivo avaliar o potencial anticoagulante de polissacarídeos sulfatados extraídos de algas de Penicillus capitatus. Os polissacarídeos sulfatados extraídos foram purificados, parcialmente caracterizados e sua atividade anticoagulante foi avaliada. O processo de extração seguido pela precipitação com etanol resultou em cinco frações. Entre as frações analisadas, F44 foi a maior concentração de polissacarídeos sulfatados. Após a purificação, as frações F23 e F44 mostraram atividade anticoagulante in vitro em um teste de tempo de tromboplastina parcialmente ativada e tempo de protrombina. Seu mecanismo preferencial é baseado nas interações entre trombina e fator Xa. Estudos adicionais sobre a estrutura farmacológica são necessários para testar a viabilidade do uso como agente terapêutico.
RESUMO
Utilization of marine algae has increased considerably over the past decades, since biodiversity within brown, red and green marine algae offers possibilities of finding a variety of bioactive compounds. Marine algae are rich sources of dietary fibre. The remarkable positive effects of seaweed dietary fibre on human body are related to their prebiotic activity over the gastrointestinal tract (GIT) microbiota. However, dietary modulation of microorganisms present in GIT can be influenced by different factors such as type and source of the dietary fibre, their molecular weight, type of extraction and purification methods employed, composition and modification of polysaccharide and oligosaccharide. This review will demonstrate evidence that polysaccharides and oligosaccharides from marine algae can be used as prebiotics, emphasizing their use in human health, their application as food and other possible applications. Furthermore, an important approach of microbial enzymes employment during extraction, modification or production of those prebiotics is highlighted.
Assuntos
Clorófitas/metabolismo , Oligossacarídeos/metabolismo , Polissacarídeos/metabolismo , Prebióticos/análise , Alga Marinha/metabolismo , Microbioma Gastrointestinal/efeitos dos fármacos , Humanos , Oligossacarídeos/química , Oligossacarídeos/farmacologia , Phaeophyceae/metabolismo , Polissacarídeos/química , Polissacarídeos/farmacologia , Rodófitas/metabolismoRESUMO
OBJECTIVE: We investigated the effect of intermittent food restriction (IFR) cycles on hypothalamic expression of lipogenic proteins and induction of overeating. METHODS: Female Wistar rats were distributed in three groups: free access to feed (control, C), 2 d feed restriction at 50% of C intake followed by 3 d (restricted 3, R3) or 5 d (restricted 5, R5) ad libitum feeding. After 6 wk, the rats were submitted to euthanasia and collected the hypothalamus and blood. The deposits of retroperitoneal, mesenteric, and gonadal fat were weighed. The expression of the mRNA for sterol regulatory element binding protein (SREBP) 1c and 2 and acetyl-CoA carboxylase in the hypothalamus were determined by real-time polymerase chain reaction, and glucose and triacylglycerol were evaluated by a commercial kit. Body mass and food intake were measured daily. RESULTS: IFR promoted increased expression of SREBP-2 in both treated groups and, in R5, increased expression of SREBP-1c. The serum triacylglycerol, mesenteric deposit, and total fat content were higher in R3. Neither of the treatment intervals altered the expression of the mRNA of acetyl-CoA carboxylase enzyme but induced hyperglycemia and higher food intake immediately after food restriction. CONCLUSION: IFR affected the expression of SREBP-1c in R5 and SREBP-2 in the hypothalamus and caused overeating immediately after fasting in both groups. We suggest that hypothalamic and peripheral alterations, coupled with compulsive eating behavior in the ad libitum period, indicate risks for diabetes mellitus and recovery of body mass after interruption of IFR.
Assuntos
Restrição Calórica/efeitos adversos , Ingestão de Alimentos/genética , Jejum/efeitos adversos , Hiperfagia/metabolismo , Proteínas de Ligação a Elemento Regulador de Esterol/metabolismo , Acetil-CoA Carboxilase/metabolismo , Animais , Feminino , Hiperfagia/etiologia , Hipotálamo/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos WistarRESUMO
The encapsulated yeast Cryptococcus neoformans is the causative agent of cryptococosis, an opportunistic life-threatening infection. C. neoformans is coated by a polysaccharide capsule mainly composed of glucuronoxylomannan (GXM). GXM is considered a key virulence factor of this pathogen. The present work aimed at evaluating the effects of GXM on the key glycolytic enzyme, 6-phosphofructo-1-kinase (PFK). GXM inhibited PFK activity in cultured murine macrophages in both dose- and time-dependent manners, which occurred in parallel to cell viability decrease. The polysaccharide also inhibited purified PFK, promoting a decrease on the enzyme affinity for its substrates. In macrophages GXM and PFK partially co-localized, suggesting that internalized polysaccharide directly may interact with this enzyme. The mechanism of PFK inhibition involved dissociation of tetramers into weakly active dimers, as revealed by fluorescence spectroscopy. Allosteric modulators of the enzyme able to stabilize its tetrameric conformation attenuated the inhibition promoted by GXM. Altogether, our results suggest that the mechanism of GXM-induced cell death involves the inhibition of the glycolytic flux.
Assuntos
Cryptococcus neoformans/patogenicidade , Macrófagos/enzimologia , Fosfofrutoquinase-1/antagonistas & inibidores , Polissacarídeos/farmacologia , Regulação Alostérica , Animais , Morte Celular , Células Cultivadas , Regulação para Baixo/efeitos dos fármacos , Glicólise , Macrófagos/efeitos dos fármacos , Camundongos , Multimerização Proteica/efeitos dos fármacos , Estabilidade Proteica/efeitos dos fármacosRESUMO
Glycosaminoglycans (GAGs) from the integument of Lithobates catesbeianus were biochemically characterized and histochemically localized. Moreover, carbohydrate distribution was investigated using conventional and lectin histochemistry at light microscopy. Hyaluronan (HA), dermatan sulfate (DS) and a heparanoid were found in the integument. Sulfated and carboxylated GAGs were visualized in the Eberth-Katschenko (EK) layer, in the mucous glands, in the hypodermis as well as in the mast cells. Furthermore, glucose and galactose were identified in the integument through thin layer chromatography (TLC) assays. N-Acetyl-beta-glucosamine residues were identified in the mucous glandular cells, between the corneum and spinosum strata, in the subepidermal region, and in the EK layer. N-Acetyl-galactosamine residues were evident in the EK layer, corresponding to a residue of the dermatan sulfate chain, which may be related to the collagenous fiber arrangement. These glycoconjugates occurred as secretory glandular products and as dermal structural elements. Moreover, HA and DS are the predominant GAGs in the L. catesbeianus integument. Considering the importance of glycoconjugates, they play a significant role to the integrity of the skin, providing mechanical support for integument cells. In addition, they are important to the water regulation mechanisms, since L. catesbeianus is preferably aquatic.
Assuntos
Glicoconjugados/análise , Glicosaminoglicanos/análise , Ranidae , Pele/química , Animais , Cromatografia em Camada Fina , Histocitoquímica , Lectinas/metabolismo , Masculino , Microscopia , Ligação ProteicaRESUMO
The interaction between phenolic substances (PS) and alginates (ALG) has been suggested to play a role in the structure of the cell walls of brown seaweeds. However, no clear evidence for this interaction was reported. Vanadium bromoperoxidase (VBPO) has been proposed as a possible catalyst for the binding of PS to ALG. In this work, we studied the interaction between PS and ALG from brown algae using size exclusion chromatography (SEC) and optical tweezers microscopy. The analysis by SEC revealed that ALG forms a high-molecular-weight complex with PS. To study the formation of this molecular complex, we investigated the in vitro interaction of purified ALG from Fucus vesiculosus L. with purified PS from Padina gymnospora (Kütz.) Sond., in the presence or absence of VBPO. The interaction between PS and ALG only occurred when VBPO was added, indicating that the enzyme is essential for the binding process. The interaction of these molecules led to a reduction in ALG viscosity. We propose that VBPO promotes the binding of PS molecules to the ALG uronic acids residues, and we also suggest that PS are components of the brown algal cell walls.