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Integrative proteomic and microRNA analysis of the intestine of mice with Type 2 Diabetes Mellitus treated with different concentrations of fluoride in drinking water

Grant number: 19/26117-8
Support Opportunities:Scholarships in Brazil - Post-Doctorate
Effective date (Start): July 01, 2020
Effective date (End): October 31, 2023
Field of knowledge:Health Sciences - Dentistry - Social and Preventive Dentistry
Principal Investigator:Marília Afonso Rabelo Buzalaf
Grantee:Aline Dionizio Valle
Host Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil


Diabetes Mellitus (DM) is considered a disease of great impact and the prospects are that it should increase considerably in the coming years, due to people's lifestyles. Among the two types of diabetes mellitus, type 2 (DM2) is the most prevalent (90%). The pathogenesis of DM2 is very complex and is involved with a progression of insulin resistance in the liver and peripheral tissues, which is further accompanied by a defective insulin secretion by pancreatic ² cells, leading to an abnormal increase in blood glucose. Some studies have indicated that fluoride (F) may influence insulin resistance/sensitivity, an early feature of DM2 development. Thus, studies relating this pathology to the administration of F are important, since this ion is present in water supply and other sources, such as toothpaste and food. As the main route of F absorption is the Gastrointestinal Tract (GIT) and the main organ is the intestine, the aim of this study is to evaluate the proteomic and miRNA changes caused by F in the intestine of animals with DM2, induced by hyperlipidic diet and streptozotocin administration, besides implementing a new line of research (miRNA analysis) in the biochemistry laboratory of the Bauru School of Dentistry - USP. Eighteen mice adult male (C57BL/6J) will be used and divided into 3 experimental groups (n = 6) according to the dose of F (0, 10 or 50 mgF/L) administered by drink. The animals will receive a hyperlipidic diet and after reaching 30g and at least 8 weeks, will receive daily intraperitoneal streptozotocin (STZ) injections of 40mg/kg for 3 to 5 days. From the 4th week will begin treatment with F, lasting 21 days. Blood glucose will be measured weekly using a glycemic monitor (Accu-Check Performa, Roche Diagnostics, Mannheim, Germany) to detect insulin resistance, and after STZ administration and DM2 testing. A group of 6 mice (C57BL /6) will be used as control. They will receive for the same experimental period, water without F and normocaloric diet. After the experimental period, the animals will be euthanized for sample collection, and blood will be collected for F analysis (specific ion electrode), as well as the intestine for marker-free quantitative proteomic analysis (Protein Linx Global Service software) and microRNA. After verifying the normality and homoscedasticity of the data, they will be submitted to the appropriate statistical analysis (p <0.05). (AU)

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