PROTEOMIC AND HISTOLOGICAL ANALYSIS OF KIDNEYS FROM MICE WITH TYPE 2 DIABETES MELLITUS TREATED WITH DIFFERENT FLUORIDE CONCENTRATIONS IN DRINK WATER

Abstract

Diabetes mellitus (DM) is a disease of great social impact, and the prospects are that its prevalence will increase even more, due to people's lifestyle. Among the different types of diabetes, type 2 (DM2) is the one with the highest prevalence (90%). The pathogenesis of DM2 is complex, and is involved with a progression of insulin resistance in the liver and peripheral tissues, accompanied by a defective secretion of insulin by pancreatic ² cells, leading to hyperglycemia. Recent studies have indicated that fluoride (F) can influence insulin resistance/sensitivity in mice. Thus, studies that correlate DM with fluoride administration are of great importance, since F is present in public water supply and in dental products. Knowing that the kidney is the main organ responsible for F excretion/retention, the objective of this study is to evaluate the proteomic and histological alterations caused by the chronic treatment with F by two doses (10mgF/L and 50mgF/L) in the kidney of mice with DM2 induced by a high-fat diet and the administration of streptozotocin. For this purpose, 30 adult male mice of the C57BL/6J strain were used, which were divided into three experimental groups (n=10), according to the dose of F administered in the drinking water, namely: D0 (0mgF/L), D10 (10mgF/L) and D50 (50mgF/L). The animals received a high-fat diet and after reaching 30g and at least 8 weeks, they received daily injections of streptozotocin (STZ) intraperitoneally (40mg/kg) for 3 days, after one week, diabetes was confirmed (9 weeks), blood glucose e200mg/dL, treatment with F was started for 21 days (3 weeks). A group with 10 mice of the C57BL/6 strain was used as control. They underwent the same experimental period, receiving water without F and a normocaloric diet. After the experimental period, the animals were euthanized for sample collection, the right kidney being collected for histological analysis and the left kidney for proteomic analysis. For the histological analysis of the kidney, the samples will be processed and stained with hematoxylin-eosin, periodic acid-Schiff and Masson's Trichrome. For marker-free quantitative proteomic analysis (Protein Linx Global Service Software) the left kidney will be processed following a protocol established by the laboratory. After verifying the normality and homoscedasticity of the data, they will be subjected to appropriated statistical analysis (p<0.05).