Efeito da dieta hiperlipídica e melatonina sobre a lesão periapical na sensibilidade a insulina, inflamação, perfil lipídico e estresse oxidativo em musculo esquelético de ratos
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Data
2022-12-19
Autores
Santos, Rodrigo Martins dos [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Atualmente, na literatura, existem diversos tipos de dietas alimentares administradas em animais que visam verificar os efeitos em seu metabolismo e em sua saúde sistêmica, com intuito de compreender como essas dietas podem afetar a saúde humana. Sendo assim, é fundamental verificar a relação entre o consumo de dietas ricas em gordura juntamente com infecções endodônticas associadas, tais como: a periodontite apical (PA), pois ambas compartilham efeitos sistêmicos como a resistência insulínica, e o aumento de mediadores inflamatórios. A melatonina (MEL) vem sendo utilizada em cenários clínicos para várias doenças crônicas (diabetes, dislipidemias, doenças cardiovasculares), além de patologias orais; isso se deve as suas propriedades antioxidantes e anti-inflamatórias. Portanto, o objetivo deste estudo foi investigar os efeitos do tratamento com ME em ratos com periodontite apical (PA) alimentados com dieta hiperlipídica (DHL) sob à sensibilidade a insulina, citocinas plasmáticas, expressão proteica e gênica de Toll-like receptor tipo 4 (TLR-4) e sua via de sinalização e estresse oxidativo no músculo gastrocnêmico (MG). Para tanto, 80 ratos Wistar com 60 dias foram distribuídos em 8 grupos (n=10): 1) controle (CN); 2) PA; 3) DHL; 4) DHL+PA; 5) CN+MEL); 6) PA+MEL; 7) DHL+MEL; 8) DHL+PA+MEL. Os grupos com DHL foram alimentados com uma dieta composta por 45,5% de ração padrão + 22,7% de banha animal e gordura vegetal + 9% de açúcar; os demais grupos receberam dieta padrão. No 7º dia os grupos com PA foram submetidos a indução cirúrgica da PA e, após 70 dias da indução da PA, os animais do grupo com melatonina receberam MEL (5 mg/Kg) diluída na água de beber, por 30 dias. Ao término do tratamento foram avaliados os seguintes parâmetros: 1) ingestão hídrica, alimentar e evolução da massa corpórea; 2) Histologia dos maxilares; 3) glicemia, insulinemia e HOMA-IR; 4) concentrações plasmáticas de TNF-α, IL-1β, IL-6 e LPS; 5) perfil lipídico; 6) expressão gênica e proteica do TLR4 e sua via de sinalização intracelular: MyD88, TRIF, IRF-3 e NFκB no MG; 7) estresse oxidativo: proteína carbonilada (PC) e substâncias reativas ao ácido tiobarbitúrico (TBARs), capacidade antioxidante total não-enzimática (FRAP) e das concentrações de glutationa reduzida (GSH), superóxido disumutase (SOD), Catalase (CAT) e Glutationa peroxidase (GPx). A análise estatística foi feita por análise de variância ANOVA seguida pelo teste de Tukey, o nível de significância será de 5%. Os resultados do presente estudo demonstraram que: 1) Os grupos alimentados com DHL apresentaram diminuição da massa corpórea, menor ingestão hídrica, alimentar e maior ingestão energética quando comparados aos grupos alimentados com dieta padrão; 2) Os grupos DHL e DHLPA apresentaram aumento do peso absoluto do tecido adiposo periepididimal (TAB); os grupos alimentados com DHL apresentaram diminuição do peso MG; o tratamento com ME promoveu diminuição do peso do TAB; 3) Os grupos DHL e DHLPA apresentaram aumento da glicemia; o grupo DHLPA aumento da insulinemia; PA, DHL e DHLPA demonstraram resistência insulínica e o tratamento com MEL reverteu as alterações na glicemia, insulinemia e resistência insulínica 4) Perfil lipídico: os grupos DHL e DHLPA apresentaram aumento de LDL e diminuição de HDL, o tratamento com ME reverteu esta alteração; 5) Citocinas: os grupos PA, DHL e DHLPA mostraram aumento do TNF-α, e a ME reverteu; o tratamento com MEL diminuiu os níveis de IL-1β plasmáticos nos grupos PAME e DHLMEL; não houve alterações nos níveis de IL-6 plasmáticos; houve redução na concentração plasmática de LPS apenas no grupo DHLME; 6) Expressão gênica e proteica: houve aumento da expressão proteica de IRF-3 nos grupos PA, DHL e DHLPA no MG e o tratamento com MEL diminuiu a expressão da proteína apenas nos grupos PAMEL e DHLMEL; 7) Não houve alterações na expressão gênica do TLR4 e sua via de sinalização celular; 8) Estresse oxidativo: TBARS aumentou nos grupos PA e DHLPA, e MEL reverteu apenas no grupo DHLPAMEL; PC aumentou em todos os grupos DHL e melhorou nos grupos MEL; a atividade da GSH diminuiu em todos os grupos experimentais, e a MEL aumentou esse parâmetro apenas nos grupos CNME e PAME; FRAP não se alterou entre os grupos, mas o tratamento com ME aumentou sua atividade nos grupos PAMEL e DHLMEL; a MEL aumentou a SOD nos grupos CN e AP. Conclui-se que a associação da DHL e da PA intensificou as alterações observadas neste estudo. O tratamento com ME demonstrou ser um importante recurso para reversão destas alterações devido aos seus efeitos antioxidantes, anti-inflamatório e hipocolesterolêmicos. Sendo assim estudos clínicos devem ser realizados para verificar os efeitos sistêmicos e inflamatórios do PA e DHL.
Currently in the literature there are several types of diets administered to animals that aim to verify their effects on metabolism and systemic health of animals, in order to understand how these diets can affect human health. Therefore, it is essential to verify the relationship between the consumption of high-fat diets together with associated endodontic infections such as apical periodontitis (AP), since both share systemic effects such as insulin resistance and increased inflammatory mediators. Currently, melatonin (MEL) has been used in clinical scenarios for several chronic diseases (diabetes, dyslipidemia, cardiovascular diseases) in addition to oral pathologies; all this thanks to its antioxidant and anti-inflammatory properties. Therefore, the aim of this study was to investigate the effects of ME treatment in rats with apical periodontitis (AP) fed a high-fat diet (HFD) on insulin sensitivity, plasma cytokines, protein and gene expression of Toll-like receptor type 4 (TLR-4) and it signaling pathway and oxidative stress in the gastrocnemius muscle (GM). For this purpose, 80 Wistar rats aged 60 days were divided into 8 groups (n=10): 1) control (CN); 2) AP; 3) HFD; 4) HFD+AP; 5) CN+MEL); 6) AP+MEL; 7) HFD+MEL; 8) HFD+AP+MEL. HFD groups were fed a diet consisting of 45.5% standard chow + 22.7% animal fat and vegetable fat + 9% sugar; the other groups received a standard diet. On the 7th day, the groups with BP underwent surgical induction of BP and, 70 days after AP induction, the animals in the melatonin group received MEL (5 mg/Kg) diluted in drinking water for 30 days. At the end of treatment, the following parameters were evaluated: 1) water and food intake and evolution of body mass; 2) Histology of the jaws; 3) glycemia, insulinemia and HOMA-IR; 4) plasma concentrations of TNF-α, IL-1β, IL-6 and LPS; 5) lipid profile; 6) gene and protein expression of TLR4 and its intracellular signaling pathway: MyD88, TRIF, IRF-3 and NFκB in MG; 7) oxidative stress: protein carbonyl (PC) and thiobarbituric acid reactive substances (TBARs), total non-enzymatic antioxidant capacity (FRAP) and concentrations of reduced glutathione (GSH), superoxide disumutase (SOD), Catalase (CAT) and Glutathione peroxidase (GPx). Statistical analysis was performed by analysis of variance ANOVA followed by Tukey's test, the significance level will be 5%. The results of the present study demonstrated that: 1) The groups fed HFD showed a decrease in body mass, lower water and food intake and higher energy intake when compared to the groups fed a standard diet; 2) The HFD and HFDAP groups showed an increase in the absolute weight of the periepididymal adipose tissue (WAT); groups fed with DHL showed a decrease in MG weight; treatment with ME promoted a decrease in WAT weight; 3) The HFD and DHLPA groups showed increased glycemia; the HFDAP group increased insulinemia; AP, HFD and HFDAP showed insulin resistance and treatment with MEL reversed changes in glycemia, insulinemia and insulin resistance 4) Lipid profile: HFD and HFDAP groups showed an increase in LDL and a decrease in HDL, treatment with ME reversed this change; 5) Cytokines: the AP, HFD and HFDAP groups showed an increase in TNF-α, and ME was reversed; treatment with MEL decreased plasma IL-1β levels in the APMEL and HFDMEL groups; there were no changes in plasma IL-6 levels; there was a reduction in the plasma concentration of LPS only in the HFDMEL group; 6) Gene and protein expression: there was an increase in the protein expression of IRF-3 in the AP, HFD and HFDAP groups in MG and the treatment with MEL decreased the protein expression only in the APMEL and HFDMEL groups; 7) There were no changes in TLR4 gene expression and its cell signaling pathway; 8) Oxidative stress: TBARS increased in the AP and HFDPA groups, and MEL reversed only in the HFDAPMEL group; CP increased in all DHL groups and improved in MEL groups; GSH activity decreased in all experimental groups, and MEL increased this parameter only in CNMEL and APME groups; FRAP did not change between groups, but ME treatment increased its activity in APMEL and HFDMEL groups; MEL increased SOD in the CN and AP groups. It is concluded that the association of HFD and AP intensified the alterations observed in this study. ME treatment proved to be an important resource for reversing these changes due to its antioxidant, anti-inflammatory and hypocholesterolemic effects. Therefore, clinical studies must be carried out to verify the systemic and inflammatory effects of AP and HFD.
Currently in the literature there are several types of diets administered to animals that aim to verify their effects on metabolism and systemic health of animals, in order to understand how these diets can affect human health. Therefore, it is essential to verify the relationship between the consumption of high-fat diets together with associated endodontic infections such as apical periodontitis (AP), since both share systemic effects such as insulin resistance and increased inflammatory mediators. Currently, melatonin (MEL) has been used in clinical scenarios for several chronic diseases (diabetes, dyslipidemia, cardiovascular diseases) in addition to oral pathologies; all this thanks to its antioxidant and anti-inflammatory properties. Therefore, the aim of this study was to investigate the effects of ME treatment in rats with apical periodontitis (AP) fed a high-fat diet (HFD) on insulin sensitivity, plasma cytokines, protein and gene expression of Toll-like receptor type 4 (TLR-4) and it signaling pathway and oxidative stress in the gastrocnemius muscle (GM). For this purpose, 80 Wistar rats aged 60 days were divided into 8 groups (n=10): 1) control (CN); 2) AP; 3) HFD; 4) HFD+AP; 5) CN+MEL); 6) AP+MEL; 7) HFD+MEL; 8) HFD+AP+MEL. HFD groups were fed a diet consisting of 45.5% standard chow + 22.7% animal fat and vegetable fat + 9% sugar; the other groups received a standard diet. On the 7th day, the groups with BP underwent surgical induction of BP and, 70 days after AP induction, the animals in the melatonin group received MEL (5 mg/Kg) diluted in drinking water for 30 days. At the end of treatment, the following parameters were evaluated: 1) water and food intake and evolution of body mass; 2) Histology of the jaws; 3) glycemia, insulinemia and HOMA-IR; 4) plasma concentrations of TNF-α, IL-1β, IL-6 and LPS; 5) lipid profile; 6) gene and protein expression of TLR4 and its intracellular signaling pathway: MyD88, TRIF, IRF-3 and NFκB in MG; 7) oxidative stress: protein carbonyl (PC) and thiobarbituric acid reactive substances (TBARs), total non-enzymatic antioxidant capacity (FRAP) and concentrations of reduced glutathione (GSH), superoxide disumutase (SOD), Catalase (CAT) and Glutathione peroxidase (GPx). Statistical analysis was performed by analysis of variance ANOVA followed by Tukey's test, the significance level will be 5%. The results of the present study demonstrated that: 1) The groups fed HFD showed a decrease in body mass, lower water and food intake and higher energy intake when compared to the groups fed a standard diet; 2) The HFD and HFDAP groups showed an increase in the absolute weight of the periepididymal adipose tissue (WAT); groups fed with DHL showed a decrease in MG weight; treatment with ME promoted a decrease in WAT weight; 3) The HFD and DHLPA groups showed increased glycemia; the HFDAP group increased insulinemia; AP, HFD and HFDAP showed insulin resistance and treatment with MEL reversed changes in glycemia, insulinemia and insulin resistance 4) Lipid profile: HFD and HFDAP groups showed an increase in LDL and a decrease in HDL, treatment with ME reversed this change; 5) Cytokines: the AP, HFD and HFDAP groups showed an increase in TNF-α, and ME was reversed; treatment with MEL decreased plasma IL-1β levels in the APMEL and HFDMEL groups; there were no changes in plasma IL-6 levels; there was a reduction in the plasma concentration of LPS only in the HFDMEL group; 6) Gene and protein expression: there was an increase in the protein expression of IRF-3 in the AP, HFD and HFDAP groups in MG and the treatment with MEL decreased the protein expression only in the APMEL and HFDMEL groups; 7) There were no changes in TLR4 gene expression and its cell signaling pathway; 8) Oxidative stress: TBARS increased in the AP and HFDPA groups, and MEL reversed only in the HFDAPMEL group; CP increased in all DHL groups and improved in MEL groups; GSH activity decreased in all experimental groups, and MEL increased this parameter only in CNMEL and APME groups; FRAP did not change between groups, but ME treatment increased its activity in APMEL and HFDMEL groups; MEL increased SOD in the CN and AP groups. It is concluded that the association of HFD and AP intensified the alterations observed in this study. ME treatment proved to be an important resource for reversing these changes due to its antioxidant, anti-inflammatory and hypocholesterolemic effects. Therefore, clinical studies must be carried out to verify the systemic and inflammatory effects of AP and HFD.
Descrição
Palavras-chave
Melatonina, Resistência à insulina, Estresse oxidativo, Doenças periodontais, Receptor 4 toll-like, Inflamação, Inflammation