GENE EXPRESSION ANALYSIS OF ENZYMES AND TRANSPORTERS RELATED TO CARNOSINE METABOLISM IN SKELETAL MUSCLE TISSUE (SOLEUS) OF SPRAGUE-DAWLEY RATS FED A HYPERLIPIDEMIC DIET AFTER BETA-ALANINE SUPPLEMENTATION

• Grant number: 23/14144-6
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Effective date (Start): October 01, 2024
• Effective date (End): September 30, 2025
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Ana Carolina Magalhães
• Grantee: Evilyn Parães Fernandes
• Host Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil

Abstract

The Metabolic Syndrome (MS), responsible for Oxidative Stress (OS) in body metabolism, is related to the development of many chronic non-communicable diseases, such as diabetes mellitus and systemic arterial hypertension, making clinical treatments that reduce OS in the body relevant. Consequently, studies that demonstrate the effectiveness of beta-alanine (²A) supplementation as an antioxidant in combating free radicals and reactive oxygen species are significant. Therefore, the objective of this study is to evaluate, based on the gene expression of enzymes and transporters related to carnosine metabolism in skeletal muscle (soleus) tissue, the effect of ²A supplementation in Sprague-Dawley rats (300-400g) fed a hyperlipidemic diet. The animals will be divided into 4 groups of 10, totalizing 40 rats: 10 with a standard diet only (CO) (without MS + CO), 10 with a standard diet and ²A supplementation (CO+²A) (without MS + CO with ²A supplementation), 10 with MS and a hyperlipidic diet (MS), and 10 with MS, ²A supplementation, and a hyperlipidic diet (MS+²A). The study period will take 10 weeks, during which the animals will have free access to food and water. Starting from the 4th week, the CO+²A and MS+²A groups will receive a dose of 250 mg/kg/day of ²A through gavage, while the MS and CO groups will receive a placebo. Blood concentrations of triglycerides, glucose, insulin, total cholesterol, leptin, and glucose tolerance will be analyzed. Subsequently, skeletal striated muscle (soleus) will be collected for RT-PCR. Regarding the RT-PCR analysis, total RNA will be extracted using TRIzol, followed by reverse transcription into cDNA using the QuatiTect Reverse Transcription® kit. The resulting mRNA will be analyzed through quantitative PCR reactions using the Taqman® system. This method will be repeated to evaluate the expression of 5 genes related to carnosine. All values will be presented as mean ± standard error of the mean (SEM). Parametric tests will be used for samples with a normal distribution, and non-parametric tests will be applied to samples with non-normal distribution using GraphPad Prism software (USA), considering p<0.05.