O efeito da ingestão de hesperidina sobre a modulação da resposta inflamatória e nível de perda óssea em camundongos com doença periodontal induzida
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Data
2020-03-30
Autores
Ramadan, Dania
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Universidade Estadual Paulista (Unesp)
Resumo
O objetivo do estudo foi avaliar o efeito da hesperidina(Hesp) sobre a periodontite experimental induzida por injeção de LPS de E. coli em camundongos. 50 camundongos BALB/c foram submetidos a diferentes tratamentos: dieta convencional, dieta enriquecida com hesperidina (25 e 50 mg/kg de peso corporal/dia) e dieta com naproxeno 10mg/kg de peso corporal/dia. Após 30 dias de suplementação os animais foram submetidos a periodontite induzida pela injeção bilateral de Escherichia coli 3x/semana por 4 semanas. Avaliou-se o peso corporal, consumo alimentar e energético ao longo de todo período experimental. Depois de 3 dias da última injeção de LPS os animais foram eutanasiados, coletados fígado e maxila e realizada a análise microscópica do fígado, quantificação de perda óssea por micro-Ct, análise microscópica e estereométrica dos maxilares, perfil de citocinas pró e anti-inflamatórias por imunoensaio multiplex e avaliação da atividade de mieloperoxidase (MPO). In vitro foi avaliada a viabilidade celular por MTT e avaliação da proliferação celular através do método de AlamarBlue®, em cultura de fibroblastos L929 em 1,3 e 7 dias. Ao final do experimento os animais que consumiram ração enriquecida com hesperidina ou naproxeno apresentaram ganho de peso e consumo alimentar e energético semelhante ao grupo controle. Na análise microscópica do fígado não foram encontradas alterações hepáticas. O micro-CT revelou significante aumento da perda óssea no grupo LPS, no qual foi prevenida pela hesperidina ou naproxeno. A análise estereométrica revelou menor infiltrado de células inflamatórias na mucosa palatina da região de primeiro molar superior direito nos grupos que foram suplementados com hesperidina ou naproxeno. Observou-se que a suplementação alimentar com hesperidina 50mg/kg de peso corporal/dia atenuou a expressão de IL-1β. As duas concentrações de hesperidina ou naproxeno não alteraram a expressão deTNF-α quando comparadas ao grupo LPS enquanto que, a suplementação com hesperidina em ambas concentrações aumentaram a expressão de IL-10. A suplementação alimentar com hesperidina ou naproxeno reduziram a atividade de MPO com clara atividade anti-inflamatória. Os resultados in vitro não mostram efeitos citotóxicos nas concentrações mais baixas. A hesperidina não mostrou atividade metabólica de fibroblastos L929 após 7 dias de tratamento. Os resultados permitem concluir que a suplementação com hesperidina em sua dose mais alta reduziu perda óssea e infiltrado inflamatório na doença periodontal experimental sugerindo, portanto que, a hesperidina apresente ação promissora na modulação do processo inflamatório associado a doença periodontal experimental.
The aim of the study was to evaluate the effect of hesperidin(Hesp) on experimental periodontitis induced by injection of E. coli LPS in mice. 50 BALB /c mice were submitted to different treatments: conventional diet, enriched diet with hesperidin (25 and 50 mg / kg of body weight / day) and diet with naproxen 10 mg / kg of body weight / day. After 30 days of supplementation, the animals were submitted to periodontitis induced by bilateral injection of Escherichia coli 3x / week for 4 weeks. Body weight, food and energy consumption were evaluated throughout the experimental period. After 3 days of the last LPS injection, the animals were euthanized, liver and maxilla were collected and microscopic analysis of the liver was carried out, quantification of bone loss by micro-Ct, microscopic and stereometric analysis of the jaws, pro and anti-inflammatory cytokine profile by multiplex immunoassay and evaluation of myeloperoxidase (MPO) activity. In vitro cell viability was evaluated by MTT and evaluation of cell proliferation using the AlamarBlue method, in culture of L929 fibroblasts in 1.3 and 7 days. At the end of the experiment, the animals that consumed enriched food with hesperidin or naproxen showed weight gain and food and energy consumption similar to the control group. Microscopic analysis of the liver showed no liver abnormalities. The micro-CT revealed a significant increase in bone loss in the LPS group, in which it was prevented by hesperidin or naproxen. The stereometric analysis revealed a lower infiltration of inflammatory cells in the palatal mucosa of the upper right first molar region in groups that were supplemented with hesperidin or naproxen. It was observed that food supplementation with hesperidin 50mg / kg of body weight / day attenuated the expression of IL-1β. The two concentrations of hesperidin or naproxen did not alter the expression of TNF-α when compared to the LPS group, whereas supplementation with hesperidin in both concentrations increased the expression of IL-10. Food supplementation with hesperidin or naproxen reduced the activity of MPO with clear anti-inflammatory activity. In vitro results do not show cytotoxic effects at lower concentrations. Hesperidin did not show metabolic activity of L929 fibroblasts after 7 days of treatment. The results allow us to conclude that supplementation with hesperidin in its highest dose reduced bone loss and inflammatory infiltrate in experimental periodontal disease, suggesting, therefore, that hesperidin presents a promising action in modulating the inflammatory process associated with experimental periodontal disease.
The aim of the study was to evaluate the effect of hesperidin(Hesp) on experimental periodontitis induced by injection of E. coli LPS in mice. 50 BALB /c mice were submitted to different treatments: conventional diet, enriched diet with hesperidin (25 and 50 mg / kg of body weight / day) and diet with naproxen 10 mg / kg of body weight / day. After 30 days of supplementation, the animals were submitted to periodontitis induced by bilateral injection of Escherichia coli 3x / week for 4 weeks. Body weight, food and energy consumption were evaluated throughout the experimental period. After 3 days of the last LPS injection, the animals were euthanized, liver and maxilla were collected and microscopic analysis of the liver was carried out, quantification of bone loss by micro-Ct, microscopic and stereometric analysis of the jaws, pro and anti-inflammatory cytokine profile by multiplex immunoassay and evaluation of myeloperoxidase (MPO) activity. In vitro cell viability was evaluated by MTT and evaluation of cell proliferation using the AlamarBlue method, in culture of L929 fibroblasts in 1.3 and 7 days. At the end of the experiment, the animals that consumed enriched food with hesperidin or naproxen showed weight gain and food and energy consumption similar to the control group. Microscopic analysis of the liver showed no liver abnormalities. The micro-CT revealed a significant increase in bone loss in the LPS group, in which it was prevented by hesperidin or naproxen. The stereometric analysis revealed a lower infiltration of inflammatory cells in the palatal mucosa of the upper right first molar region in groups that were supplemented with hesperidin or naproxen. It was observed that food supplementation with hesperidin 50mg / kg of body weight / day attenuated the expression of IL-1β. The two concentrations of hesperidin or naproxen did not alter the expression of TNF-α when compared to the LPS group, whereas supplementation with hesperidin in both concentrations increased the expression of IL-10. Food supplementation with hesperidin or naproxen reduced the activity of MPO with clear anti-inflammatory activity. In vitro results do not show cytotoxic effects at lower concentrations. Hesperidin did not show metabolic activity of L929 fibroblasts after 7 days of treatment. The results allow us to conclude that supplementation with hesperidin in its highest dose reduced bone loss and inflammatory infiltrate in experimental periodontal disease, suggesting, therefore, that hesperidin presents a promising action in modulating the inflammatory process associated with experimental periodontal disease.
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Palavras-chave
Flavonoides, Doenças Periodontais, Hesperidina, Anti-Inflamatórios, Flavonoids, Periodontal diseases, Hesperidin, Anti-Inflammatory Agents