RESUMO
Fucoidans are sulfated polysaccharides that are found in marine algae and have many useful activities, including antitumor effects, promotion of apoptosis of cancer cells, and antiviral, anti-inflammatory, and antiallergic actions. In oral medicine, several case reports have shown that fucoidan-containing creams and tablets markedly improved recurrent aphthous stomatitis, symptomatic inflamed tongue, and recurrent oral herpes labialis. The aim of this study was to examine the properties of fucoidans for use in oral healthcare. The antimicrobial, anti-adhesion, endotoxin-neutralizing, and cyclooxygenase (COX)-1 and COX-2 inhibitory activities of fucoidans were examined. Four key results were obtained: fucoidans showed strong antimicrobial activity against Candida albicans, Streptococcus mutans, and Porphyromonas gingivalis; significantly inhibited the adhesion of S. mutans to bovine teeth and porcelain; were suggested to bind to and neutralize endotoxin (lipopolysaccharide) in an LAL assay; and showed COX-1 and/or COX-2 inhibitory activity. These results suggested that fucoidans may be useful in the field of oral healthcare.
Assuntos
Anti-Infecciosos , Polissacarídeos , Animais , Bovinos , Porphyromonas gingivalis , Streptococcus mutansRESUMO
Previously, we showed that the antimicrobial cationic and amphipathic octadecapeptide AmyI-1-18 from rice α-amylase (AmyI-1) inhibited the endotoxic activity of lipopolysaccharide (LPS) from Escherichia coli. In addition, we demonstrated that several AmyI-1-18 analogs containing arginine or leucine substitutions, which were designed on the basis of the helical wheel projection of AmyI-1-18, exhibited higher antimicrobial activity against human pathogenic microorganisms than AmyI-1-18. In the present study, anti-inflammatory (anti-endotoxic) activities of five AmyI-1-18 analogs containing arginine or leucine substitutions were investigated. Two single arginine-substituted and two single leucine-substituted AmyI-1-18 analogs inhibited the production of LPS-induced nitric oxide in mouse macrophages (RAW264) more effectively than AmyI-1-18. These data indicate that enhanced cationic and hydrophobic properties of AmyI-1-18 are associated with improved anti-endotoxic activity. In subsequent chromogenic Limulus amebocyte lysate assays, 50% inhibitory concentrations (IC50 ) of the three AmyI-1-18 analogs (G12R, D15R, and E9L) were 0.11-0.13 µm, indicating higher anti-endotoxic activity than that of AmyI-1-18 (IC50, 0.22 µm), and specific LPS binding activity. In agreement, surface plasmon resonance analyses confirmed direct LPS binding of three AmyI-1-18 analogs. In addition, AmyI-1-18 analogs exhibited little or no cytotoxic activity against RAW264 cells, indicating that enhancements of anti-inflammatory and LPS-neutralizing activities following replacement of arginine or leucine did not result in significant increases in cytotoxicity. This study shows that the arginine-substituted and leucine-substituted AmyI-1-18 analogs with improved anti-endotoxic and antimicrobial activities have clinical potential as dual-function host defense agents. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Peptídeos Catiônicos Antimicrobianos/farmacologia , Arginina/química , Leucina/química , Lipopolissacarídeos/antagonistas & inibidores , Proteínas de Plantas/farmacologia , alfa-Amilases/farmacologia , Sequência de Aminoácidos , Substituição de Aminoácidos , Animais , Anti-Inflamatórios não Esteroides/síntese química , Anti-Inflamatórios não Esteroides/química , Peptídeos Catiônicos Antimicrobianos/síntese química , Peptídeos Catiônicos Antimicrobianos/química , Linhagem Celular , Humanos , Interações Hidrofóbicas e Hidrofílicas , Teste do Limulus , Lipopolissacarídeos/química , Lipopolissacarídeos/farmacologia , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Óxido Nítrico/antagonistas & inibidores , Óxido Nítrico/biossíntese , Oryza/química , Proteínas de Plantas/síntese química , Proteínas de Plantas/química , Ligação Proteica , Relação Estrutura-Atividade , alfa-Amilases/síntese química , alfa-Amilases/químicaRESUMO
Food proteins have been identified as a source of bioactive peptides. These peptides are inactive within the sequence of the parent protein and must be released during gastrointestinal digestion, fermentation, or food processing. Of bioactive peptides, multifunctional cationic peptides are more useful than other peptides that have specific activity in promotion of health and/or the treatment of diseases. We have identified and characterized cationic peptides from rice enzymes and proteins that possess multiple functions, including antimicrobial, endotoxin-neutralizing, arginine gingipain-inhibitory, and/or angiogenic activities. In particular, we have elucidated the contribution of cationic amino acids (arginine and lysine) in the peptides to their bioactivities. Further, we have discussed the critical parameters, particularly proteinase preparations and fractionation or purification, in the enzymatic hydrolysis process for producing bioactive peptides from food proteins. Using an ampholyte-free isoelectric focusing (autofocusing) technique as a tool for fractionation, we successfully prepared fractions containing cationic peptides with multiple functions.
Assuntos
Anti-Infecciosos/química , Peptídeos Catiônicos Antimicrobianos/química , Proteínas de Plantas/química , Anti-Infecciosos/uso terapêutico , Peptídeos Catiônicos Antimicrobianos/uso terapêutico , Arginina/química , Fermentação , Manipulação de Alimentos , Humanos , Hidrólise , Focalização Isoelétrica , Proteínas do Leite/química , Oryza/químicaRESUMO
We have previously reported that AmyI-1-18, an octadecapeptide derived from α-amylase (AmyI-1) of rice, is a novel cationic α-helical peptide that exhibited antimicrobial activity against human pathogens, including Porphyromonas gingivalis, Pseudomonas aeruginosa, Propionibacterium acnes, Streptococcus mutans, and Candida albicans. In this study, to further investigate the potential functions of AmyI-1-18, we examined its inhibitory ability against the endotoxic activities of lipopolysaccharides (LPSs, smooth and Rc types) and lipid A from Escherichia coli. AmyI-1-18 inhibited the production of endotoxin-induced nitric oxide (NO), an inflammatory mediator, in mouse macrophages (RAW264) in a concentration-dependent manner. The results of a chromogenic Limulus amebocyte lysate assay illustrated that the ability [50% effective concentration (EC50): 0.17 µM] of AmyI-1-18 to neutralize lipid A was similar to its ability (EC50: 0.26 µM) to neutralize LPS, suggesting that AmyI-1-18 specifically binds to the lipid A moiety of LPS. Surface plasmon resonance analysis of the interaction between AmyI-1-18 and LPS or lipid A also suggested that AmyI-1-18 directly binds to the lipid A moiety of LPS because the dissociation constant (KD) of AmyI-1-18 with lipid A is 5.6×10(-10) M, which is similar to that (4.3×10(-10) M) of AmyI-1-18 with LPS. In addition, AmyI-1-18 could block the binding of LPS-binding protein to LPS, although its ability was less than that of polymyxin B. These results suggest that AmyI-1-18 expressing antimicrobial and endotoxin-neutralizing activities is useful as a safe and potent host defense peptide against pathogenic Gram-negative bacteria in many fields of healthcare.