RESUMO
Fucogalactomannan (FGM) is a non-sulphated polysaccharide isolated from the Tylopilus ballouii mushroom. We investigated the chemical characteristics of this FGM using HPLC, chemical methods, and NMR studies ((1)H, (13)C, (1)H/(13)C-HSQC and DEPT-135 spectroscopies) without chemical fragmentation. This polysaccharide consisted primarily of mannose and galactose with variable amounts of fucose and traces of xylose and with MW of 140kDa. Infrared and NMR spectroscopies showed the possible interaction between these polysaccharides and proteins. The antioxidant activity showed for FGM a high inhibition of superoxide and hydroxyl radicals with an IC50 of 1.25 and 1.6mg/mL, respectively. The results of peroxidation tests showed that FGM had an IC50 of 1.72mg/mL. Furthermore, the anti-inflammatory assay showed that FGM reduced edema by 32.8%, 42.0%, and 56% at doses of 30, 50, and 70mg/kg, respectively. Thus, these results suggested a structure and indicated possible anti-inflammatory and antioxidant activities use of these polysaccharides.
Assuntos
Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Antioxidantes/química , Antioxidantes/farmacologia , Basidiomycota/química , Polissacarídeos/química , Polissacarídeos/farmacologia , Animais , Anti-Inflamatórios/isolamento & purificação , Anti-Inflamatórios/uso terapêutico , Antioxidantes/isolamento & purificação , Antioxidantes/uso terapêutico , Edema/tratamento farmacológico , Edema/patologia , Camundongos , Peso Molecular , Monossacarídeos/análise , Polissacarídeos/isolamento & purificação , Polissacarídeos/uso terapêuticoRESUMO
BACKGROUND: Red and brown algae sulfated polysaccharides (SPs) have been widely investigated as antinociceptive and/or anti-inflammatory agents; however, no description of these biological properties concerning green algae SPs have been reported. Caulerpa curpressoides (Chlorophyta) presents three SPs fractions (Cc-SP1, Cc-SP2, and Cc-SP3). Anticoagulant (in vitro) and anti- and pro-thrombotic (in vivo) effects of Cc-SP2 had been recently reported. We evaluated the effects of Cc-SP2 using models of nociception and acute inflammation in vivo. METHODS: Male Swiss mice received Cc-SP2 (iv) 30 min prior to receiving 0.6% acetic acid (10 ml/kg, ip), 1% formalin (20 µl, sc) or were subjected to thermal stimuli (51 ± 1 °C). Cc-SP2 was injected sc to male Wistar rats in a peritonitis model or a paw edema model using carrageenan (ip or ipl, 500 µg). To analyze the systemic effects, Cc-SP2 (27 mg/kg, sc) was administrated to both genders mice before waiting for 14 days. RESULTS: Cc-SP2 (3, 9 or 27 mg/kg) reduced (p < 0.05) the number of writhes induced by acetic acid by 57, 89.9 and 90.6%, respectively, the licking time in the first (9 or 27 mg/kg with 42.47 and 52.1%, respectively) and the second (3, 9 or 27 mg/kg with 68.95, 82.34 and 84.61%, respectively) phases. In the hot-plate test, the antinociceptive effect of Cc-SP2 (9 mg/kg) was primarily observed at 60 min (26.7 ± 1.2 s), with its effect reversed by naloxone (8.6 ± 1.3 s), suggesting the involvement of the opioid system. Cc-SP2 (3, 9 or 27 mg/kg, sc, p < 0.05) showed anti-inflammatory effects by decreasing neutrophils migration by 64, 69 and 73%, respectively, and potently reduced the paw edema, especially at the second (0.16 ± 0.02, 0.16 ± 0.03 and 0.12 ± 0.05 ml) and third (0.16 ± 0.03, 0.18 ± 0.02 and 0.14 ± 0.04 ml) hours, respectively. Cc-SP2 did not cause hepatic or renal alterations or affect body mass or the macroscopy of the organs examined (p > 0.05). Histopathological analyses of the liver and kidney showed that both organs were affected by Cc-SP2 treatment, but these effects were considered reversible. CONCLUSION: The results indicate that the analgesic and anti-inflammatory effects of Cc-SP2 could be of biomedical applicability as a new, natural tool in pain and acute inflammatory conditions.
Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Caulerpa/química , Clorófitas/química , Edema/tratamento farmacológico , Dor/tratamento farmacológico , Peritonite/tratamento farmacológico , Polissacarídeos/uso terapêutico , Doença Aguda , Animais , Anti-Inflamatórios não Esteroides/efeitos adversos , Anti-Inflamatórios não Esteroides/isolamento & purificação , Modelos Animais de Doenças , Feminino , Coração/efeitos dos fármacos , Rim/efeitos dos fármacos , Rim/patologia , Fígado/efeitos dos fármacos , Fígado/patologia , Masculino , Camundongos , Miocárdio/patologia , Medição da Dor , Polissacarídeos/efeitos adversos , Polissacarídeos/isolamento & purificação , Ratos , Ratos WistarRESUMO
A sulfated polysaccharide (SPSG) was successfully isolated from seagrass Halodule wrightii Asch., Cymodoceaceae, and its antioxidant and anticoagulant activities were investigated. The data presented here showed that the SPSG is a 11 kDa sulfated heterogalactan with a sulfatation degree of 20.63 percent and it also contains glucose and xylose. SPSG antioxidant activities were evaluated using several in vitro assays and the anticoagulant activity was evaluated by aPTT and PT tests. These assays suggested that the SPSG possessed remarkable antioxidant properties in different in vitro assays and an outstanding anticoagulant activity 2.5-fold higher than that of heparin Clexane® in the aPTT test. This data represents the first reported on the sulfated polysaccharide biological activities from seagrass. These results indicate that SPSG can be considered in the future as a drug utilized in treating diseases from these systems.
RESUMO
Fucan is a term used to denominate a family of sulfated polysaccharides rich in L-fucose. The brown alga Spatoglossum schröederi, Dictyotaceae, synthesizes three heterofucans named A, B, and C. Fucan A is a non-anticoagulant heterofucan which possesses potent antithrombotic (in vivo) and antiproliferative (in vitro) activities. However, its toxicity in vivo has not been determined. The present study examined the acute and subchronic toxicity of the fucan A in Wistar rats after subcutaneous administration. After that, the animals were killed and examined. The results showed in the acute study that fucan A did not cause general adverse effects and mortality in the concentrations 0, 20, 100, 1000, and 2000 µg/g body weight per rat for seven days. Regarding the subchronic study, the data showed that the fucan A did not cause any change in hematological and biochemistry parameters, as well as in the morphology, and in the size of the rat's organs analyzed at a concentration of 20 µg/g body weight per rat during a 62-day period. In conclusion, this study indicates this heterofucan is a compound with potential pharmacological value that has no toxicity in vivo.
RESUMO
The anti-inflammatory properties of a heparin-like compound from the shrimp Litopenaeus vannamei are related. Besides reducing significantly (p<0.001) the influx of inflammatory cells to injury site in a model of acute inflammation, shrimp heparin-like compound was able to reduce the matrix metalloproteinase (MMPs) activity in the peritoneal lavage of inflamed animals. Moreover, this compound also reduced almost 90% the activity of MMP-9 secreted by human activated leukocytes. Negligible anti-coagulant activities in aPPT assay and a poor bleeding potential make this compound a better alternative than mammalian heparin as a possible anti-inflammatory drug.
Assuntos
Anti-Inflamatórios/farmacologia , Anticoagulantes/farmacologia , Glicosaminoglicanos/farmacologia , Heparina/farmacologia , Inflamação/tratamento farmacológico , Penaeidae/fisiologia , Animais , Anti-Inflamatórios/química , Anticoagulantes/química , Endotélio Vascular/citologia , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/metabolismo , Glicosaminoglicanos/química , Glicosaminoglicanos/isolamento & purificação , Hemorragia/tratamento farmacológico , Heparina/química , Heparina/isolamento & purificação , Heparitina Sulfato/metabolismo , Leucócitos/efeitos dos fármacos , Leucócitos/enzimologia , Metaloproteinase 9 da Matriz/metabolismo , Neutrófilos/efeitos dos fármacos , Cavidade Peritoneal/fisiologia , Coelhos , Ratos , SuínosRESUMO
Fucan is a term used to denominate a family of sulfated L-fucose-rich polysaccharides. The brown alga Spatoglossum schröederi (Dictyotaceae) has three heterofucans namely fucan A, B and C. The 21 kDa fucan A is composed of a core of a beta (1-3) glucuronic acid-containing oligosaccharide of 4.5 kDa with branches at C4 of the fucose chains alpha (1-3) linked. The fucose is mostly substituted at C4 with a sulfate group and at C2 with chains of beta (1-4) xylose. This fucan has neither anticoagulant (from from 0.1 to 100 microg) nor hemorrhagic activities (from 50 to 800 microg/mL). The antithrombotic test in vivo showed that fucan A has no activity in any of the concentrations (from 0.2 to 20 microg/g/day) tested 1 h after polysaccharide administration. However, when fucan A was injected endovenously 24 h before the ligature of the venae cavae, we observed a dose-dependent effect, reaching saturation at around 20 microg/g of rat weight. In addition, this effect is also time-dependent, reaching saturation around 16 h after fucan administration. In addition, regardless of the administration route, fucan A displayed antithrombotic activity. The exception was the oral pathway. Of particular importance was the finding that fucan A stimulates the synthesis of an antithrombotic heparan sulfate from endothelial cells like heparin. The hypothesis has been raised that the in vivo antithrombotic activity of fucan A is related to the increased production of this heparan. Taken together with the fact that the compound is practically devoid of anticoagulant and hemorrhagic activity, the data suggest that it may be an ideal antithrombotic agent in vivo.
Assuntos
Células Endoteliais/efeitos dos fármacos , Fibrinolíticos/isolamento & purificação , Heparitina Sulfato/biossíntese , Phaeophyceae/química , Polissacarídeos/isolamento & purificação , Animais , Linhagem Celular , Relação Dose-Resposta a Droga , Células Endoteliais/metabolismo , Fibrinolíticos/administração & dosagem , Humanos , Masculino , Polissacarídeos/efeitos adversos , Coelhos , Ratos , Ratos Wistar , Fatores de TempoRESUMO
The Geastrum saccatum a mushroom, native to Brazil, is produced under natural conditions in the unexplored reserve of Mata da Estrela-RN. This species has curative properties for eye infections and diseases such as asthma. The tissues of this mushroom contain carbohydrates, proteins, lipids, moisture and ashes in amounts of 42.3%, 37.05%, 9.01, 1.4% and 10.2%, respectively. An extract from this mushroom was characterized by chemical analyses and (13)C and (1)H NMR spectroscopy. It contains high amount of glucose and traces of galactose. The signal appearing at 103.5 ppm was assigned to C1 of beta-glucose. The signals observed between 20 and 40 ppm suggest the presence of a glucan-protein compound. This glucan inhibited the lipid peroxidation at the dose of 0.27 mg/mL (59.1%) and it can protect cells against oxidative stress by scavenging of the hydroxyl (77%) and superoxide (88.4%) radicals at 0.27 mg/mL. The glucan (30 mg/kg) reduces the polymorphonuclear cell migration (57.6%). The ear edema induced by croton oil was inhibited by glucan (60.4% at 10 mg/kg) and by its association with diclofenac (5 mg/kg) (89.2%) or L-NAME (60 mg/kg) (86.23%). Histological analyses of the ear edema induced by croton oil in the presence of glucan (10, 30 or 50 mg/kg) showed a reduced degree of the polymorphonuclear cell migration. We concluded that the glucan has antioxidant, and antiinflammatory properties as well as its antiinflammatory effect are mediated by inhibition of both nitric oxide synthase (NOS) and cyclooxygenase (COX).
Assuntos
Agaricales , Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Produtos Biológicos/farmacologia , Citotoxinas/farmacologia , beta-Glucanas/farmacologia , Animais , Edema/tratamento farmacológico , Proteínas Fúngicas/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Pleurisia/tratamento farmacológico , Ratos , Ratos WistarRESUMO
The brown alga Spatoglossum schroederi contains three fractions of sulfated polysaccharides. One of them was purified by acetone fractionation, ion exchange, and molecular sieving chromatography. It has a molecular size of 21.5 kDa and contains fucose, xylose, galactose, and sulfate in a molar ratio of 1.0:0.5:2.0:2.0 and contains trace amounts of glucuronic acid. Chemical analyses, methylation studies, and NMR spectroscopy showed that the polysaccharide has a unique structure, composed of a central core formed mainly by 4-linked beta-galactose units, partially sulfated at the 3-O position. Approximately 25% of these units contain branches of oligosaccharides (mostly tetrasaccharides) composed of 3-sulfated, 4-linked alpha-fucose and one or two nonsulfated, 4-linked beta-xylose units at the reducing and nonreducing end, respectively. This sulfated galactofucan showed no anticoagulant activity on several "in vitro" assays. Nevertheless, it had a potent antithrombotic activity on an animal model of experimental venous thrombosis. This effect is time-dependent, reaching the maximum 8 h after its administration compared with the more transient action of heparin. The effect was not observed with the desulfated molecule. Furthermore, the sulfated galactofucan was 2-fold more potent than heparin in stimulating the synthesis of an antithrombotic heparan sulfate by endothelial cells. Again, this action was also abolished by desulfation of the polysaccharide. Because this sulfated galactofucan has no anticoagulant activity but strongly stimulates the synthesis of heparan sulfate by endothelial cells, we suggested that this last effect may be related to the "in vivo" antithrombotic activity of this polysaccharide. In this case the highly sulfated heparan sulfate produced by the endothelial cells is in fact the antithrombotic agent. Our results suggested that this sulfated galactofucan may have a potential application as an antithrombotic drug.
Assuntos
Fibrinolíticos/farmacologia , Fucose/química , Hemostasia , Phaeophyceae/metabolismo , Acetona/química , Acetona/farmacologia , Animais , Anticoagulantes/química , Anticoagulantes/farmacologia , Sequência de Carboidratos , Bovinos , Cromatografia , Cromatografia por Troca Iônica , Meios de Cultivo Condicionados/farmacologia , Relação Dose-Resposta a Droga , Eletroforese em Gel de Ágar , Células Endoteliais/citologia , Células Endoteliais/metabolismo , Fator Xa/química , Fibrinolíticos/química , Furanos/química , Galactose/química , Ácido Glucurônico/química , Heparina/química , Heparitina Sulfato/química , Humanos , Espectroscopia de Ressonância Magnética , Metilação , Dados de Sequência Molecular , Oligossacarídeos/química , Polissacarídeos/química , Ratos , Enxofre/química , Ésteres do Ácido Sulfúrico/química , Timidina/química , Fatores de Tempo , Xilose/químicaRESUMO
In recent years, sulfated fucans have emerged as an important class of natural biopolymers. In this study, the anti-adhesive activity of a fucan from the brown seaweed Spatoglossum schröederi was analyzed using tumorigenic cells: wild-type Chinese hamster ovary cells (CHO-K1) and the mutant type deficient in xylosyltransferase (CHO-745). Fibronectin (FN) was used as substrate for cell attachment. For both cell types, this fucan has shown a dose-dependent anti-adhesive effect, reaching saturation at around 400 mug/mL. This effect was abolished by desulfation of the fucan. In addition, this polymer exhibited the highest inhibitory effect in comparison to other sulfated polysaccharides. The fucan was biotinylated and used as a probe to identify its action sites. Biotinylated fucan was detected in the extracellular matrix environment by confocal microscopy and flow cytometric analysis, but not at the cell surface. The results suggest that the fucan shows anti-adhesive activity by binding directly to FN, and blocking FN sites that are recognized by cell surface ligands, possibly the integrin family.
Assuntos
Anticoagulantes/farmacologia , Adesão Celular/efeitos dos fármacos , Fibronectinas , Fitoterapia , Polissacarídeos/farmacologia , Alga Marinha , Animais , Anticoagulantes/administração & dosagem , Anticoagulantes/uso terapêutico , Células CHO/efeitos dos fármacos , Cricetinae , Cricetulus , Dissacarídeos , Relação Dose-Resposta a Droga , Matriz Extracelular/química , Feminino , Citometria de Fluxo , Extratos Vegetais/administração & dosagem , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Polissacarídeos/administração & dosagem , Polissacarídeos/uso terapêuticoRESUMO
Alpha-amylase Inhibitors were isolated from Ficus sp. (Gameleira) seeds by acetone fractionation and Sephadex G-50. Two inhibitors (alpha-PPAI and alpha-ZSAI) were tested against alpha-amylases from coleopteran larvae. alpha-PPAI was active to alpha-amylases of Callosobruchus maculatus (52%) and Zabrotes subfasciatus (53%). alpha-ZSAI was strongly active to Z. subfasciatus (100%) of and Mimosestes mimosae (98%). The alpha-ZSAI is a glycoprotein of approximately 50 kDa with an IC50 value of 0.074 microg microl(-1).