RESUMO
A large number of infections are caused by Gram-positive and Gram-negative multi-resistant bacteria worldwide, adding up to a figure of around 700,000 deaths per year. The indiscriminate uses of antibiotics, as well as their misuse, resulted in the selection of bacteria resistant to known antibiotics, for which it has little or no treatment. In this way, the strategies to combat the resistance of microorganisms are extremely important and, essential oils of Croton species have been extensively studied for this purpose. The aim of this study was to carry the evaluation of antibacterial, antibiofilm, antioxidant activities, and spectroscopic investigation of essential oil from Croton piauhiensis (EOCp). The EOCp exhibited antimicrobial activity against Gram-positive and Gram-negative bacteria with required MICs ranging from 0.15 to 5% (v/v). In addition, the MBC of the EOCp for Staphylococcus aureus ATCC 25923 and ATCC 700698, were 0.15 and 1.25%, respectively. Moreover, the EOCp significantly reduced significantly the biofilm production and the number of viable cells from the biofilm of all bacterial strains tested. The antioxidant potential of the EOCp showed EC50 values ranging from 171.21 to 4623.83 µg/mL. The EOCp caused hemolysis (>45%) at the higher concentrations tested (1.25 to 5%), and minor hemolysis (17.6%) at a concentration of 0.07%. In addition, docking studies indicated D-limonene as a phytochemical with potential for antimicrobial activity. This study indicated that the EOCp may be a potential agent against infections caused by bacterial biofilms, and act as a protective agent against ROS and oxidative stress.
Assuntos
Anti-Infecciosos , Croton , Óleos Voláteis , Antibacterianos/farmacologia , Antioxidantes/farmacologia , Biofilmes , Bactérias Gram-Negativas , Bactérias Gram-Positivas , Testes de Sensibilidade Microbiana , Óleos Voláteis/farmacologiaRESUMO
Schinus terebinthifolius Raddi (Anacardiaceae) is popularly known in Brazil as aroeira-da-praia and has pharmacological use as an astringent, antidiarrheal, anti-inflammatory, depurative, diuretic, and antifebrile agent. Although the neuropathic antinociceptive potential of S. terebinthifolius fruits has already been investigated, this study is the first one to analyze the acute antinociceptive effect of the essential oil of S. terebinthifolius (female) leaves (EOFSt) on adult zebrafish. EOFSt was submitted to antioxidant activity evaluation by two methods (ferrous ion-chelating capacity [FIC] and ß-carotene). The animals (n = 6/group) were treated orally (20 µL) with EOFSt (0.1, 0.5, or 1.0 mg/mL) or vehicle (0.9% sodium chloride [NaCl]; 20 µL), and submitted to nociception (formalin, cinnamaldehyde, capsaicin, glutamate, acidic saline, and hypertonic saline). Possible neuromodulation mechanisms, as well motor alterations and toxicity were also evaluated. In the FIC assay, EOFSt showed ferrous ion-chelating capacity in â¼40% to 90%. Regarding the ß-carotene bleaching assay, EOFSt showed inhibition in a 58% to 80% range. Oral administration of EOFSt showed no acute toxicity and did not alter the locomotor system of aZF, and reduced the nociceptive behavior in all tested models. These effects of EOFSt were significantly similar to those of morphine, used as a positive control. The antinociceptive effect of EOFSt was inhibited by naloxone, L-NAME, ketamine, camphor, ruthenium red, and amiloride. The antinociceptive effect of the EOFSt cornea was inhibited by capsazepine. EOFSt has the pharmacological potential for acute pain treatment and this effect is modulated by the opioid system, NMDA receptors, and transient receptor potential ankyrin 1 (TRPA1), transient receptor potential vanilloid 1 (TRPV1), and acid-sensing ion channels. The EOFSt also has the pharmacological potential for corneal pain treatment and this effect is modulated by the TRPV1 channel.
Assuntos
Anacardiaceae/química , Analgésicos/farmacologia , Óleos Voláteis/farmacologia , Peixe-Zebra/fisiologia , Administração Oral , Analgésicos/química , Animais , Anti-Inflamatórios não Esteroides/química , Anti-Inflamatórios não Esteroides/farmacologia , Antioxidantes/metabolismo , Feminino , Masculino , Óleos Voláteis/química , Folhas de Planta/químicaRESUMO
This study aimed to determinate the chemical composition and evaluate the antimicrobial and antioxidant activity of the essential oil obtained from leaves of V. gardneriana. The Vitex gardneriana leaves's were hydrodistilled to obtain the essential oil and the chemical composition determined by GC/MS analysis. The antimicrobial activities were determined by microdilution method. The activity of essential oil on biofilm was evaluated by quantification of total biomass and enumeration of biofilm-entrapped viable cells. The antioxidant activity was assessed by DPPH free radical assay, ferrous ion chelating assay, ferric-reducing antioxidant power and ß-carotene bleaching assay. Furthermore, the essential oil was tested on viability of health human, animal cells and the microcrustacean Artemia sp. The essential oil showed high content of sesquiterpenes and very low content of monoterpenes. Regarding activity on planktonic cells, the essential oil reduced the growth of the all species tested but showed MIC values only to S. aureus (0.31%). In general, the essential oil reduced significantly the biofilm biomass and the number of viable cells of bacteria and yeasts, mainly on biofilm formation. The essential oil showed a potential antioxidant activity, mainly on ß-carotene oxidation. Moreover, the essential oil reduced the cell viability of murine fibroblasts but not show viability reduction of human keratinocytes. Furthermore, the oil not show toxicity against the microcrustacean. Thus, the essential oil from V. gardneriana leaves may be considered as an important alternative against biofilms formed by bacteria and yeasts related to infections, as well as a natural antioxidant and non-toxic substance on human cells.
Assuntos
Anti-Infecciosos/química , Antioxidantes/química , Óleos Voláteis/química , Extratos Vegetais/química , Folhas de Planta/química , Vitex/química , Animais , Anti-Infecciosos/farmacologia , Artemia/efeitos dos fármacos , Bactérias/efeitos dos fármacos , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Brasil , Candida/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Camundongos , Testes de Sensibilidade Microbiana , Monoterpenos/química , Óleos Voláteis/isolamento & purificação , Óleos Voláteis/farmacologia , Extratos Vegetais/farmacologia , Pseudomonas aeruginosa/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , beta CarotenoRESUMO
This study evaluated the antibacterial, antifungal, and antioxidant effect of 7-hydroxy-4',6-dimethoxy-isoflavone and essential oil of Myroxylon peruiferum. The compound was isolated and its structure elucidated by NMR. The chemical composition of essential oil determined by GC-MS analysis. To evaluation of antimicrobial activity, the Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) and Minimum Fungicidal Concentration (MFC) were performed. In addition to analysis of antioxidant activity, DPPH radical scavenging tests, iron chelating assay (FIC), antioxidant reducing power assay (FRAP) and ß-carotene bleaching assay (BCB) were performed. For the essential oil were identified 24 organized compounds having as main constituents; Germacrene D (17.2%), α-pinene (14.8%) and E-caryophyllene (10.8%). The results showed that isoflavone (2000 to 156 µg/mL) and essential oil (5.0 to 1.25%) present antibacterial and antifungal activity against Gram-positive bacteria and filamentous fungi. The isoflavone and the essential oil also presented antioxidant activity in all the tests, mainly on inhibition of the oxidation of ß-carotene test concentrations ranging from 60 to 100%. In conclusion, isoflavone and essential oil from M. peruiferum present an antimicrobial alternative against Gram-positive bacteria, especially of the genus Staphylococcus and dermatophyte fungi of the genus Trichophyton, as well as a natural compound antioxidant.
Assuntos
Antibacterianos/farmacologia , Antifúngicos/farmacologia , Myroxylon/química , Óleos Voláteis/farmacologia , Extratos Vegetais/farmacologia , Antioxidantes/análise , Bactérias/efeitos dos fármacos , Fungos/efeitos dos fármacos , Cromatografia Gasosa-Espectrometria de Massas , Quelantes de Ferro , Isoflavonas/análise , Testes de Sensibilidade Microbiana , Óleos Voláteis/química , Extratos Vegetais/química , Valores de Referência , Reprodutibilidade dos Testes , Fatores de TempoRESUMO
ABSTRACT This study evaluated the chemical composition and antioxidant activity of fatty acids from the marine red algae Pterocladiella capillacea (S. G. Gmelin) Santelices & Hommersand 1997 and Osmundaria obtusiloba (C. Agardh) R. E. Norris 1991. The gas chromatography mass spectrometry (GC-MS) identified nine fatty acids in the two species. The major fatty acids of P. capillacea and O. obtusiloba were palmitic acid, oleic acid, arachidonic acid and eicosapentaenoic acid. The DPPH radical scavenging capacity of fatty acids was moderate ranging from 25.90% to 29.97%. Fatty acids from P. capillacea (31.18%) had a moderate ferrous ions chelating activity (FIC), while in O. obtusiloba (17.17%), was weak. The ferric reducing antioxidant power (FRAP) of fatty acids from P. capillacea and O. obtusiloba was low. As for β-carotene bleaching (BCB), P. capillacea and O. obtusiloba showed a good activity. This is the first report of the antioxidant activities of fatty acids from the marine red algae P. capillacea and O. obtusiloba.
Assuntos
Rodófitas/química , Ácidos Graxos/análise , Ácidos Graxos/química , Antioxidantes/análise , Antioxidantes/química , Valores de Referência , Análise de Variância , Sequestradores de Radicais Livres/análise , beta Caroteno/análise , FMN Redutase/análise , Cromatografia Gasosa-Espectrometria de MassasRESUMO
This study evaluated the chemical composition and antioxidant activity of fatty acids from the marine red algae Pterocladiella capillacea (S. G. Gmelin) Santelices & Hommersand 1997 and Osmundaria obtusiloba (C. Agardh) R. E. Norris 1991. The gas chromatography mass spectrometry (GC-MS) identified nine fatty acids in the two species. The major fatty acids of P. capillacea and O. obtusiloba were palmitic acid, oleic acid, arachidonic acid and eicosapentaenoic acid. The DPPH radical scavenging capacity of fatty acids was moderate ranging from 25.90% to 29.97%. Fatty acids from P. capillacea (31.18%) had a moderate ferrous ions chelating activity (FIC), while in O. obtusiloba (17.17%), was weak. The ferric reducing antioxidant power (FRAP) of fatty acids from P. capillacea and O. obtusiloba was low. As for ß-carotene bleaching (BCB), P. capillacea and O. obtusiloba showed a good activity. This is the first report of the antioxidant activities of fatty acids from the marine red algae P. capillacea and O. obtusiloba.
Assuntos
Antioxidantes/análise , Antioxidantes/química , Ácidos Graxos/análise , Ácidos Graxos/química , Rodófitas/química , Análise de Variância , FMN Redutase/análise , Sequestradores de Radicais Livres/análise , Cromatografia Gasosa-Espectrometria de Massas , Valores de Referência , Fatores de Tempo , beta Caroteno/análiseRESUMO
OBJECTIVE: To evaluate the antioxidant, antibacterial and bacterial cell agglutination activities of the hexane (Hex) and 70% ethanol (70% EtOH) extracts of two species of red seaweeds Pterocladiella capillacea (P. capillacea) and Osmundaria obtusiloba. METHODS: In vitro antioxidant activity was determined by DPPH radical scavenging assay, ferric-reducing antioxidant power assay, ferrous ion chelating assay, ß-carotene bleaching assay and total phenolic content quantification. Antimicrobial activity was tested using the method of disc diffusion on Mueller-Hinton medium. The ability of algal extracts to agglutinate bacterial cells was also tested. RESULTS: The 70% EtOH extract of the two algae showed the highest values of total phenolic content compared to the Hex extract. The results of DPPH for both extracts (Hex, 70% EtOH) of Osmundaria obtusiloba (43.46% and 99.47%) were higher than those of P. capillacea (33.04% and 40.81%) at a concentration of 1000 µg/mL. As for the ferrous ion chelating, there was an opposite behavior, extracts of P. capillacea had a higher activity. The extracts showed a low ferric-reducing antioxidant power, with optical density ranging from 0.054 to 0.180. Antioxidant activities of all extracts evaluated for ß-carotene bleaching were above 40%. There was no antibacterial activity against bacterial strains tested. However, the extracts of both species were able to agglutinate bacterial Gram positive cells of Staphylococcus aureus and Gram negative cells of Escherichia coli, multidrug-resistant Salmonella and Vibrio harveyi. CONCLUSIONS: This is the first report of the interaction between these algal extracts, rich in natural compounds with antioxidant potential, and Gram positive and Gram negative bacterial cells.
RESUMO
Twenty species of marine invertebrates from the Brazilian coast were screened for hemagglutinating/hemolytic activity. In at least twelve tested species, hemagglutinating activity was different for different blood types, suggesting the presence of lectins. Extracts from four species showed hemolytic activity. Two new lectins were purified from the marine sponge Cliona varians (CvL-2) and sea cucumber Holothuria grisea (HGL). CvL-2 was able to agglutinate rabbit erythrocytes and was inhibited by galactosides. The hemagglutinating activity was optimal in pH neutral and temperatures below 70 °C. CvL-2 is a trimeric protein with subunits of 175 kDa. On the other hand, HGL showed both hemagglutinating and hemolytic activity in human and rabbit erythrocytes, but hemolysis could be inhibited by osmotic protection, and agglutination was inhibited by mucin. HGL was stable in pH values ranging from 4 to 10 and temperatures up to 90 °C. In electrophoresis and gel filtration, HGL was a monomeric protein with 15 kDa. CvL-2 and HGL showed different levels of toxicity to Artemia naplii. CvL-2 showed LC50 of 850.1 µg/mL, whereas HGL showed LC50 of 9.5 µg/mL.
Assuntos
Eritrócitos/efeitos dos fármacos , Hemaglutinação/efeitos dos fármacos , Hemólise/efeitos dos fármacos , Lectinas/farmacologia , Poríferos/química , Pepinos-do-Mar/química , Animais , Artemia/efeitos dos fármacos , Brasil , Testes de Hemaglutinação , Humanos , Lectinas/classificação , Lectinas/isolamento & purificação , Poríferos/classificação , Coelhos , Pepinos-do-Mar/classificaçãoRESUMO
Marine invertebrates are capable of synthesizing bioactive compounds, which may be beneficial to human health. The aim of this study was to evaluate the antioxidant, hemolytic, antimicrobial and cytotoxic activities of crude extract (70% EtOH), and dichloromethane (DCM), ethyl acetate (EtOAc), and aqueous (Aq) fractions of the marine zoanthid Palythoa caribaeorum. The phenolic compound contents of the crude extract, DCM, EtOAc and Aq fractions were 12.33, 18.17, 10.53, and 3.18 mg GAE per gram, respectively. DPPH radical scavenging activity showed slight variation. IC50 of crude extract, DCM, EtOAc and Aq fractions were 11.13, 11.25, 11.74, and 11.28 µg mL(-1), respectively. Among the sample, ferrous ion chelating was the highest in crude extract (IC50 302.90 µg mL(-1)), followed by EtOAc, Aq, and DCM fractions with 457.77, 547.91, and 641.82 µg mL(-1), respectively. Ferric-reducing antioxidant power showed optical density at about 0.5. The samples tested exhibited low hemolytic activity under 10% up to a concentration of 50 µg mL(-1). No antimicrobial activity was observed against any of the tested bacterial strains. For the cytotoxic activity, LC50 of DCM, crude extract, EtOAc, and Aq were 52.10, 83.06, 86.34, and 117.45 µg mL(-1), showing high toxicity.
Assuntos
Antozoários/química , Antibacterianos/farmacologia , Antioxidantes/farmacologia , Artemia/efeitos dos fármacos , Hemólise/efeitos dos fármacos , Animais , Antibacterianos/isolamento & purificação , Antioxidantes/isolamento & purificação , Bioensaio , Relação Dose-Resposta a Droga , Concentração Inibidora 50 , Testes de Sensibilidade MicrobianaRESUMO
A new chromophore-containing agglutinin (Haliclona manglaris agglutinin (HMA)) was isolated from the tropical sponge H. manglaris. HMA was purified by a combination of hydrophobic interaction chromatography and ion exchange chromatography. Native HMA is a heterotrimer formed by two ß-chains (15 kDa) and one α-chain (22 kDa). HMA is a glycoprotein and possesses three intrachain disulfide bonds. Hemagglutinating activity of HMA was stable at neutral pH and temperatures up to 60 °C. HMA was only inhibited by thyroglobulin. Mass spectrometry sequencing and Edman degradation revealed a unique amino acid sequence of about 30%. Moreover, HMA has an organic chromophore of 581 Da, and this characteristic seems to be important to its antioxidant activity. Interestingly, while HMA showed no toxicity against Artemia nauplii and was unable to agglutinate bacterial cells, it did show a high capacity to protect ß-carotene against oxidation. Thus, our findings suggest the putative involvement of HMA in the protection of the sponge against oxidation.
Assuntos
Aglutininas/química , Aglutininas/isolamento & purificação , Corantes Fluorescentes/química , Haliclona/química , Sequência de Aminoácidos , Animais , Antioxidantes/farmacologia , Artemia/efeitos dos fármacos , Carboidratos/análise , Cátions Bivalentes/farmacologia , Cromatografia em Gel , Cromatografia de Fase Reversa , Eletroforese em Gel de Poliacrilamida , Hemaglutinação/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Dados de Sequência Molecular , Peso Molecular , Peptídeos/química , Coelhos , Análise de Sequência de Proteína , Compostos de Sulfidrila/química , Espectrometria de Massas em Tandem , TemperaturaRESUMO
Resveratrol can also inhibit the activation of proinflammatory mediators and cytokines at the early gene expression stage. It is well known that lectins are sugar-binding proteins that act as both pro- and anti-inflammatory molecules. Thus, the objective of this work was to verify the binding of a polyphenol compound with a lectin of Canavalia maritima (ConM) based on their ability to inhibit pro-inflammatory processes. To accomplish this, ConM was purified and crystallized, and resveratrol was soaked at 5mM for 2h of incubation. The crystal belongs to the monoclinic space group C2, the final refinement resulted in an Rfactor of 16.0% and an Rfree of 25.5%. Resveratrol binds in the rigid ß-sheet through H-bonds and hydrophobic interaction with amino acids that compose the fifth and sixth ß-strands of the rigid ß-sheet of ConM. The ConM and resveratrol inhibited DPPH oxidation, showing synergic activity with the most effective ratio of 2:3 and carbohydrate binding site is not directly related to antioxidant activity. It is the interaction between ConM and resveratrol that indicates the synergism of these two molecules in acting as free radicals scavengers and in reducing the inflammatory process through the inhibition of many pro-inflammatory events.
Assuntos
Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Lectinas de Plantas/química , Lectinas de Plantas/metabolismo , Polifenóis/química , Polifenóis/metabolismo , Estilbenos/química , Estilbenos/metabolismo , Sítios de Ligação , Compostos de Bifenilo/química , Canavalia , Cristalografia por Raios X , Sequestradores de Radicais Livres/farmacologia , Glicosilação/efeitos dos fármacos , Ligação de Hidrogênio/efeitos dos fármacos , Interações Hidrofóbicas e Hidrofílicas/efeitos dos fármacos , Simulação de Acoplamento Molecular , Picratos/química , Ligação Proteica/efeitos dos fármacos , Estrutura Secundária de Proteína , Quercetina/farmacologia , ResveratrolRESUMO
This study evaluated the effect of derriobtusone A, a flavonoid isolated from Lonchocarpus obtusus, on two important pathogenic bacteria, Staphylococcus aureus and Escherichia coli, as well as its antioxidant activity and toxicity. Planktonic growth assays were performed, and the inhibition of biofilm formation was evaluated. In addition, antioxidant activity was assessed by DPPH radical scavenging assay, ferrous ion chelating assay, ferric-reducing antioxidant power assay, and ß -carotene bleaching assay. Toxicity was evaluated by the brine shrimp lethality test. Results showed that derriobtusone A completely inhibited the planktonic growth of S. aureus at 250 and 500 µ g/mL; however, it did not have the same activity on E. coli. Derriobtusone A reduced the biomass and colony-forming unit (cfu) of S. aureus biofilm at concentrations of 250 and 500 µ g/mL. In various concentrations, it reduced the biofilm biomass of E. coli, and, in all concentrations, it weakly reduced the cfu. Derriobtusone A showed highly efficient antioxidant ability in scavenging DPPH radical and inhibiting ß -carotene oxidation. The compound showed no lethality to Artemia sp. nauplii. In conclusion, derriobtusone A may be an effective molecule against S. aureus and its biofilm, as well as a potential antioxidant compound with no toxicity.
Assuntos
Antibacterianos/administração & dosagem , Antioxidantes/administração & dosagem , Biofilmes/efeitos dos fármacos , Flavonoides/isolamento & purificação , Extratos Vegetais/administração & dosagem , Antibacterianos/química , Antioxidantes/química , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Escherichia coli/patogenicidade , Flavonoides/administração & dosagem , Flavonoides/química , Flavonoides/farmacologia , Humanos , Testes de Sensibilidade Microbiana , Oxirredução , Extratos Vegetais/química , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus aureus/patogenicidadeRESUMO
Natural antioxidants found in marine macroalgae are bioactive compounds known to play an important role in the prevention of diseases associated with aging cells protecting them against the oxidative damage. The purpose of this study was to evaluate the antioxidant and cytotoxic activity of ethanolic extracts of two species of red seaweeds, Amansia multifida and Meristiella echinocarpa. In vitro antioxidant activity was determined by DPPH radical scavenging assay, ferric-reducing antioxidant power (FRAP) assay, ferrous ion chelating (FIC) assay, ß-carotene bleaching (BCB) assay and total phenolic content (TPC) quantification. Cytotoxicity was evaluated with the brine shrimp Artemia sp. lethality test. The TPC values observed in the present study indicated that both species A. multifida and M. echinocarpa are rich in phenolic compounds, reaching values of 45.40 and 28.46 mg gallic acid equivalent (GAE) g-1 of ethanolic extract, respectively. DPPH radical scavenging and ferrous ion chelating showed values of 60% and 17%, respectively. Both seaweed extracts inhibited ß-carotene oxidation by approximately 40%. None of the algal extracts were potentially cytotoxic. The results have showed that extracts of both species of marine red algae exhibit antioxidant potential and low toxicity. They are sources of natural antioxidant compounds.
Assuntos
Antioxidantes/farmacologia , Alga Marinha/química , Animais , Antioxidantes/toxicidade , Artemia/efeitos dos fármacos , Bioensaio , Brasil , OxirreduçãoRESUMO
A novel lectin, HGA-2, was isolated from the sea cucumber Holothuria grisea. The protein was isolated by a single chromatographic step using a column of Guar Gum as affinity. HGA-2 showed an apparent molecular mass of 17 kDa and 34 kDa under reducing and nonreducing conditions, respectively. The hemagglutinating activity was specific for rabbit erythrocytes, showing no activity for human blood A, B and O. Its hemagglutinating activity was inhibited by carbohydrates containing galactose, with higher affinity for GalNAc and glycoprotein porcine stomach mucin (PSM). HGA-2 was stable at pH 6-10, significantly declining at pH 5 and a temperature of 40°C, with its activity being abolished at 100 °C. The HGA-2 protein was found to be Ca(2+)-dependent; it was highly toxic against Artemia nauplii and able to recognize and agglutinate cells of Escherichia coli. Amino acid sequences of tryptic peptides of HGA-2 strongly suggest that HGA-2 is a member of the C-type lectin family.
Assuntos
Aglutininas/química , Aglutininas/metabolismo , Escherichia coli/metabolismo , Galactosídeos/metabolismo , Holothuria/química , Lectinas/química , Lectinas/metabolismo , Aglutininas/isolamento & purificação , Aglutininas/toxicidade , Sequência de Aminoácidos , Animais , Hemaglutinação , Testes de Hemaglutinação , Humanos , Concentração de Íons de Hidrogênio , Íons , Lectinas/isolamento & purificação , Lectinas/toxicidade , Lectinas Tipo C , Dados de Sequência Molecular , Coelhos , Alinhamento de Sequência , TemperaturaRESUMO
Two new lectins named Halilectin 1 (H-1) and Halilectin 2 (H-2) were isolated from the marine sponge Haliclona caerulea using a combination of affinity chromatography on stroma fixed onto Sephadex G-25 and cation and anion exchange chromatography. H-1 is a monomeric protein with a molecular mass of 40 kDa estimated using sodium dodecyl sulfate polyacrylamide gel electrophoresis and 15 kDa estimated using a TSK gel. Conversely, H-2 is a homodimeric protein with 15 kDa monomers linked via weak interactions. H-1 more effectively agglutinates trypsinized rabbit erythrocytes, whereas H-2 more effectively agglutinates native rabbit erythrocytes. The hemagglutinating activity of H-1 could be not inhibited by any tested sugars, but H-2 was inhibited by orosomucoid and porcine stomach mucin. Neither lectin was dependent on divalent ions. H-1 was stable at basic pH range and temperatures up to 50 °C, whereas H-2 was stable at acid pH range and temperatures up to 80 °C. The H. caerulea lectins exhibited dose-dependent toxicity against Artemia nauplii. Additionally, 76% of the primary structure of H-2 was determined using tandem mass spectrometry to contain a unique amino acid sequence with no similarity to any members of the animal lectin family.
Assuntos
Haliclona/química , Lectinas/química , Lectinas/farmacologia , Poríferos/química , Sequência de Aminoácidos , Animais , Artemia/efeitos dos fármacos , Sequência de Bases , Cromatografia/métodos , Eritrócitos/efeitos dos fármacos , Hemaglutinação/efeitos dos fármacos , Testes de Hemaglutinação/métodos , Concentração de Íons de Hidrogênio , Dados de Sequência Molecular , Peso Molecular , Coelhos , Espectrometria de Massas em Tandem/métodos , TemperaturaRESUMO
Lectins are sugar-binding proteins widely distributed in nature with many biological functions. Although many lectins have a remarkable biotechnological potential, some of them can be cytotoxic. Thus, the aim of this study was to assess the toxicity of five lectins, purified from seeds of different species of Canavalia genus. In order to determine the toxicity, assays with Artemia nauplii were performed. In addition, a fluorescence assay was carried out to evaluate the binding of lectins to Artemia nauplii. In order to verify the relationship between the structure of lectins and their cytotoxic effect, structural analysis was carried out to evaluate the volume of the carbohydrate recognition domain (CRD) of each lectin. The results showed that all lectins exhibited different toxicities and bound to a similar area in the digestive tract of Artemia nauplii. Concerning the structural analysis, differences in spatial arrangement and volume of CRD may explain the variation of the toxicity exhibited by each lectin. To this date, this is the first study that establishes a link between toxicity and structure of CRD from Diocleinae lectins.
Assuntos
Artemia/efeitos dos fármacos , Artemia/metabolismo , Lectinas/toxicidade , Sequência de Aminoácidos , Animais , Artemia/química , Canavalia/toxicidade , Carboidratos/química , Lectinas/química , Lectinas/metabolismo , Sementes/químicaRESUMO
Lectins are a structurally heterogeneous group of proteins that have specific binding sites for carbohydrates and glycoconjugates. Because of their biotechnological potential, lectins are widely used in biomedical research. The present study aimed to evaluate the healing potential of the lectin isolated from the marine red alga Bryothamnion seaforthii (BSL). The lectin was purified using ion exchange chromatography with DEAE cellulose and characterized using tandem mass spectrometry. For healing tests, skin wounds were induced in the dorsal thoracic region of mice. These animals were randomly divided into three groups and subjected to topical treatment for 12 days with BSL, bovine serum albumin and 150 mM NaCl. To evaluate the potential of each treatment, the animals were anesthetized and sacrificed on days 2, 7 and 12, respectively. The parameters evaluated included the wound area, the proportion of wound closure and the histological diagnosis. The wound closure was more effective with BSL (Postoperative Day 7 and 12) than controls. The luminal epithelium was completely restructured; the presence of collagen in the dermis and the strongly active presence of young skin annexes demonstrate the potential of treatment with BSL compared with controls. Our findings suggest that BSL has pro-healing properties and can be a potential medical process in the treatment of acute wounds.
Assuntos
Lectinas/química , Lectinas/farmacologia , Rodófitas/química , Cicatrização/efeitos dos fármacos , Sequência de Aminoácidos , Animais , Derme/efeitos dos fármacos , Masculino , Camundongos , Dados de Sequência Molecular , Isoformas de Proteínas , Análise de Sequência de ProteínaRESUMO
A cianobactéria Spirulina tem sido utilizada há séculos, tendo em vista suas propriedades nutricionais e medicinais. Neste trabalho, objetivou-se analisar os teores de β-caroteno tanto em suplementos alimentares a base de Spirulina comercializados em estabelecimentos de produtos naturais no mercado varejista, quanto em S. platensis cultivada em laboratório. Alguns pigmentos carotenóides possuem atividade de vitamina A e, dentre eles, o β-caroteno é o que apresenta maior atividade biológica. A extração de β-caroteno foi feita com metanol:água (90:10 v/v), seguida de saponificação e partição em n-hexano. As análises cromatográficas foram realizadas em coluna Waters Spherisorb S5 ODS 2 (4,6 x 250 mm), usando metanol:tetrahidrofurano (90:10, v/v) bombeado a 2 mL min-1, com registro dos cromatogramas em 450 nm. Os suplementos alimentares a base de Spirulina apresentaram baixos teores de β-caroteno. A maior concentração foi encontrada em S. platensis cultivada a 24º C com fotoperíodo de 16 h claro e 8 h escuro. Considerando sua atividade provitamínica, os teores de retinol equivalente (RE) nas microalgas analisadas no presente trabalho foram calculados a partir do β-caroteno para classificá-las como fonte excelente ou fonte útil de vitamina A.
Spirulina is a cyanobacterium that has been used for several centuries due to its nutritional and medicinal properties. This work has evaluated the contents of β-carotene both in Spirulina commercialized as food supplement, purchased from natural product shops, and in S. platensis reared under laboratory conditions. Some carotenoids exhibit pro-vitamin A activity, and β-carotene presents the greatest biological activity. β-Carotene of microalgae was extracted in 90 percent aqueous methanol. These extracts were saponified and partitioned in n-hexane. Chromatographic analyses were carried out in a Spherisorb column S5 ODS 2 (4.6 x 250 mm), with a mobile phase of methanol:tetrahydrofuran (90:10, v/v) delivered at 2 mL min-1 and chromatograms registered at 450 nm absorbance. Food supplement made with Spirulina presented low content of β-carotene. The highest content of β-carotene was detected in S. platensis cultivated under 24º C with photoperiod of 16 h light and 8 h dark. Considering its pro-vitamin activity, retinol equivalent (RE) was calculated from β-carotene to determine whether alga material could be claimed as an useful or an excellent source of the vitamins A.
RESUMO
Marine algae are a promising source of beneficial compounds for human use. Among these, pro-vitamin A carotenoids and vitamins B, C, and E stand out. The objective of this study was to investigate seasonal variation of α-tocopherol levels in 5 species of green marine algae of the Caulerpa genus. This research was carried out with both fresh and dry specimens; and, in addition, differences arising as a result of the drying process were examined. Analyses were carried out by high-performance liquid chromatography (HPLC) using an isocratic system and a reversed-phase C-18 column. The distribution of α-tocopherol throughout the year in Caulerpa genus was variable. All samples of both fresh and dried algae contained α-tocopherol, except for the dried C. racemosa from March 2006. The drying process was responsible for losses of α-tocopherol ranging from 21% to 93%.
Assuntos
Clorófitas/química , Estações do Ano , alfa-Tocoferol/análise , Carotenoides/análise , Cromatografia Líquida de Alta Pressão/métodos , Vitamina A/análise , Vitamina E/análiseRESUMO
The agglutinin from the red marine alga Hypnea cervicornis (HCA) was tested in models of nociception and inflammation. The role of carbohydrate-binding sites and the systemic toxicity were assessed. HCA (10(-1), 1, and 10 mg/kg) administered i.v. to mice inhibited writhes induced by acetic acid and, at 10 mg/kg, inhibited the second phase of the formalin test, but did not alter the response latency in the hot-plate test. HCA (1 mg/kg) administered i.v. to rats reduced carrageenan-induced paw edema at 1, 2, and 3 h after challenge, but not edema induced by dextran. The neutrophil migration induced by both N-formyl-methionyl-leucyl-phenylalanine (fMLP) and carrageenan was inhibited by HCA at 10(-1), 1, and 10 mg/kg. The combination of HCA (1 mg/kg) and its ligand mucin reversed the lectin inhibitory effect on carrageenan-induced neutrophil migration and acetic acid-induced writhes. The i.v. treatment of rats with HCA (1 mg/kg) for 7 days did not affect body mass; liver, kidney or heart wet weight; blood leukocyte counts; urea, creatinine or serum transaminase activity; or macroscopy of the organs examined. In short, H. cervicornis agglutinin showed important antinociceptive and anti-inflammatory activity via interaction with the lectin carbohydrate-binding site.