RESUMO
Periodontal diseases are bacteria-induced inflammatory disorders that lead to the destruction of the tooth-supporting tissues. Active compounds endowed with a capacity to regulate the inflammatory response are regarded as potential therapeutic agents for the treatment of periodontal diseases. The aim of this study was to characterize the anti-inflammatory properties of a polyphenolic cinnamon fraction. Chromatographic and mass spectrometry analyses of the polyphenolic composition of the cinnamon fraction revealed that phenolic acids, flavonoids (flavonols, anthocyanins, flavan-3-ols), and procyanidins make up 9.22%, 0.72%, and 10.63% of the cinnamon fraction, respectively. We used a macrophage model stimulated with lipopolysaccharides (LPS) from either Aggregatibacter actinomycetemcomitans or Escherichia coli to show that the cinnamon fraction dose-dependently reduced IL-6, IL-8, and TNF-α secretion. Evidence was brought that this inhibition of cytokine secretion may result from the ability of the fraction to prevent LPS-induced NF-κB activation. We also showed that the cinnamon fraction reduces LPS binding to monocytes, which may contribute to its anti-inflammatory properties. Lastly, using a competitor assay, it was found that the cinnamon fraction may represent a natural PPAR-γ ligand. Within the limitations of this in vitro study, the cinnamon fraction was shown to exhibit a therapeutic potential for the treatment of periodontal diseases due to its anti-inflammatory properties.
Assuntos
Anti-Inflamatórios/farmacologia , Cinnamomum/química , Macrófagos/efeitos dos fármacos , Extratos Vegetais/química , Polifenóis/análise , Aggregatibacter actinomycetemcomitans/metabolismo , Antocianinas/análise , Anti-Inflamatórios/análise , Anti-Inflamatórios/química , Linhagem Celular Tumoral , Cromatografia Líquida de Alta Pressão , Cinnamomum/metabolismo , Flavonoides/análise , Humanos , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Lipopolissacarídeos/farmacologia , Macrófagos/citologia , Macrófagos/metabolismo , Espectrometria de Massas , NF-kappa B/metabolismo , Casca de Planta/química , Casca de Planta/metabolismo , Polifenóis/química , Polifenóis/farmacologia , Proantocianidinas/análise , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Cardamom (Elettaria cardamomum) is a traditional aromatic plant for which several pharmacological properties have been associated. In this study, the antibacterial activity of two cardamom extracts (fruit and seeds), rich in volatile compounds, against major periodontal pathogens was evaluated. Moreover, the ability of the extracts to exert anti-inflammatory activity was tested. Both cardamom fruit and seed extracts exerted an antibacterial effect against Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis, and Prevotella intermedia (minimum inhibitory concentrations: 0.5% [v/v], 0.25%, 0.062%, 0.125%, respectively and minimum bactericidal concentrations: 1%, 0.25%, 0.062%, 0.25%, respectively). The cell membrane of P. gingivalis was disrupted by a treatment with cardamom extracts suggesting the bactericidal mode of action. The extracts also inhibited biofilm formation although it correlated with a growth reduction. Moreover, the cardamom extracts significantly decreased the secretion of IL-1ß, TNF-α, and IL-8 by lipopolysaccharide-stimulated macrophages. Evidence were brought that the anti-inflammatory activity may result from inhibition of the NF-κB signaling pathway. This study is the first to provide evidence that cardamom fruit and seed extracts through their antibacterial and anti-inflammatory properties may be therapeutic agents of interest against periodontal infections.
Assuntos
Antibacterianos/farmacologia , Anti-Inflamatórios/farmacologia , Elettaria/química , Extratos Vegetais/farmacologia , Antibacterianos/química , Anti-Inflamatórios/química , Biofilmes/efeitos dos fármacos , Permeabilidade da Membrana Celular , Biologia Computacional/métodos , Citocinas/metabolismo , Humanos , Lipopolissacarídeos/imunologia , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Macrófagos/metabolismo , Testes de Sensibilidade Microbiana , NF-kappa B/metabolismo , Periodontite/diagnóstico , Periodontite/tratamento farmacológico , Periodontite/microbiologia , Extratos Vegetais/químicaRESUMO
Kampo formulations used in Japan to treat a wide variety of diseases and to promote health are composed of mixtures of crude extracts from the roots, bark, leaves, and rhizomes of a number of herbs. The present study was aimed at identifying the beneficial biological properties of Daiokanzoto (TJ-84), a Kampo formulation composed of crude extracts of Rhubarb rhizomes and Glycyrrhiza roots, with a view to using it as a potential treatment for periodontal disease. Daiokanzoto dose-dependently inhibited the expression of major Porphyromonas gingivalis virulence factors involved in host colonization and tissue destruction. More specifically, Daiokanzoto reduced the expression of the fimA, hagA, rgpA, and rgpB genes, as determined by quantitative real-time PCR. The U937-3xκB-LUC monocyte cell line transfected with a luciferase reporter gene was used to evaluate the anti-inflammatory properties of Daiokanzoto. Daiokanzoto attenuated the P. gingivalis-mediated activation of the NF-κB signaling pathway. It also reduced the secretion of pro-inflammatory cytokines (IL-6 and CXCL8) by lipopolysaccharide-stimulated oral epithelial cells and gingival fibroblasts. Lastly, Daiokanzoto, dose-dependently inhibited the catalytic activity of matrix metalloproteinases (-1 and -9). In conclusion, the present study provided evidence that Daiokanzoto shows potential for treating and/or preventing periodontal disease. The ability of this Kampo formulation to act on both bacterial pathogens and the host inflammatory response, the two etiological components of periodontal disease, is of high therapeutic interest.
Assuntos
Antibacterianos/farmacologia , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Glycyrrhiza/química , Medicina Kampo/métodos , Extratos Vegetais/uso terapêutico , Raízes de Plantas/química , Porphyromonas gingivalis/efeitos dos fármacos , Rheum/química , Antibacterianos/isolamento & purificação , Anti-Inflamatórios/isolamento & purificação , Anti-Inflamatórios/farmacologia , Relação Dose-Resposta a Droga , Glycyrrhiza uralensis , Humanos , Metaloproteinase 9 da Matriz/metabolismo , NF-kappa B/metabolismo , Extratos Vegetais/farmacologia , Porphyromonas gingivalis/metabolismo , Porphyromonas gingivalis/patogenicidade , Inibidores de Proteases/isolamento & purificação , Inibidores de Proteases/farmacologia , Reação em Cadeia da Polimerase em Tempo Real , Rhus , Transdução de Sinais/efeitos dos fármacos , Células U937/efeitos dos fármacos , Células U937/metabolismo , Fatores de Virulência/biossínteseRESUMO
BACKGROUND: Solobacterium moorei is a volatile sulfide compound (VSC)-producing Gram-positive anaerobic bacterium that has been associated with halitosis. The aim of this study was to investigate the effects of green tea extract and its major constituent epigallocatechin-3-gallate (EGCG) on growth and several halitosis-related properties of S. moorei. METHODS: A microplate dilution assay was used to determine the antibacterial activity of green tea extract and EGCG against S. moorei. Their effects on bacterial cell membrane integrity were investigated by transmission electron microscopy and a fluorescence-based permeability assay. Biofilm formation was quantified by crystal violet staining. Adhesion of FITC-labeled S. moorei to oral epithelial cells was monitored by fluorometry. The modulation of ß-galactosidase gene expression in S. moorei was evaluated by quantitative RT-PCR. RESULTS: The green tea extract as well as EGCG inhibited the growth of S. moorei, with MIC values of 500 and 250 µg/ml, respectively. Transmission electron microscopy analysis and a permeabilization assay brought evidence that the bacterial cell membrane was the target of green tea polyphenols. Regarding the effects of green tea polyphenols on the S. moorei colonization properties, it was found that biofilm formation on EGCG-treated surfaces was significantly affected, and that green tea extract and EGCG can cause the eradication of pre-formed S. moorei biofilms. Moreover, both the green tea extract and EGCG were found to reduce the adherence of S. moorei to oral epithelial cells. The ß-galactosidase activity of S. moorei, which plays a key role in VSC production, was dose-dependently inhibited by green tea polyphenols. In addition, EGCG at ½ MIC significantly decreased the ß-galactosidase gene expression. CONCLUSION: Our study brought evidence to support that green tea polyphenols possess a number of properties that may contribute to reduce S. moorei-related halitosis. Therefore, these natural compounds may be of interest to be used to supplement oral healthcare products.
Assuntos
Antibacterianos/farmacologia , Camellia sinensis/química , Catequina/análogos & derivados , Bactérias Gram-Positivas/efeitos dos fármacos , Halitose/microbiologia , Extratos Vegetais/farmacologia , beta-Galactosidase/antagonistas & inibidores , Antioxidantes/farmacologia , Aderência Bacteriana/efeitos dos fármacos , Biofilmes/efeitos dos fármacos , Catequina/farmacologia , Inibidores Enzimáticos/farmacologia , Bactérias Gram-Positivas/crescimento & desenvolvimento , Bactérias Gram-Positivas/metabolismo , Humanos , Polifenóis/farmacologia , Chá/químicaRESUMO
Periodontal diseases, which are inflammatory diseases of bacterial origin affecting the tooth-supporting tissues, are characterized by inflammation and destruction of gingival connective tissue and alveolar bone, and may lead to tooth loss. The aim of the study was to investigate Rokumigan, a Kampo Japanese traditional medicine made of six different plants, for its capacity to prevent biofilm formation by Fusobacterium nucleatum, to inhibit interleukin-6 (IL-6) and interleukin-8 (IL-8) secretion by mucosal cells, and to promote wound healing in a fibroblast model. Using a microplate colorimetric assay, Rokumigan prevented biofilm formation by F. nucleatum, while it had no effect on bacterial growth. Rokumigan inhibited IL-6 secretion in both epithelial cells and fibroblasts stimulated with lipopolysaccharide. However, it caused no significant inhibition of IL-8 secretion by both cell types. Rokumigan significantly increased proliferation and migration of gingival fibroblasts in a wound healing assay. In conclusion, the Kampo formulation Rokumigan, through suppression of biofilm formation by F. nucleatum, inhibition of IL-6 secretion by gingival epithelial cells and fibroblasts, and promotion of wound healing in a fibroblast model, may have potential application for periodontal diseases.