Evaluation of protein content and DNA methylation of GLUT4 in the gastrocnemius muscle of adult rats, offspring of rats with apical periodontitis

Abstract

Fetal programming suggests that stimuli applied during early fetal development can alter the offspring's metabolism, increasing the risk of disease in adulthood. DNA methylation is one of the epigenetic mechanisms involved in the regulation of gene expression. Previous studies have found that offspring of rats with apical periodontitis (AP) present in adult life: insulin resistance, alteration in the initial step of the insulin signal (decrease in the phosphorylation of IRS1 / IRS2 status, after insulin stimulation) in the gastrocnemius muscle (GM) and increase in the plasma concentration oftumor necrosis-alpha (TNF-alpha). It is known that pro-inflammatory cytokines, such as TNF-alpha, can decrease the gene expression of GLUT4, which is a fundamental transporter in the insulin signaling pathway and involved in insulin-mediated glucose uptake in skeletal muscle. The objective of the present study is to evaluate the effects of maternal AP in rats on GLUT4 in relation to its content, its translocation to the plasma membrane and the degree of DNA methylation in the GM of its adult offspring. For that, the 18 Wistar rats (2 months old) will be distributed in 3 groups: 1) control rats; 2) rats with 1 AP induced in the 1st right upper molar; 3) rats with 4 APs induced in the 1st and 2nd molars of the upper and lower right side. The AP will be induced using a carbon steel drill with a 0.1 mm sphere at the end. After 30 days of pulp exposure, rats from all groups will be placed for mating. When the male offs ofpring all the rats are 75 days old, the following analyzes will be performed in the GM: 1) GLUT4 content and its translocation index to the plasma membrane by western blot method; 2) degree of DNA methylation in the promoter region of the GLUT4 gene by Restriction Digestion and Real-Time PCR - qAMP method. The statistical analysis will be done by ANOVA, followed by the Tukey test (p<0.05).