Autophagy involvement in the metabolic and inflammatory alterations in adipose tissue associated with Obesity induced by high fat diet

Abstract

In follow-up to the master's degree experiments (FAPESP#2016/05406-3), in this project of doctorate, we will test the hypothesis that the autophagy pathway is an important mediator of the metabolic, morphological and inflammatory alterations of adipose tissue in Obesity. During the master's, we performed an experimental protocol where wild-type C57BL6/J mice fed with control or high fat diet were treated with the pharmacological inhibitor of autophagy chloroquine (CQ) or water for 8 weeks. Briefly, we found that mice treated with CQ are partially protected from Obesity (weight gain and adiposity) and glucose intolerance induced by the ingestion of high fat diet. For the doctorate, we propose to investigate a possible participation of nonshivering thermogenesis in the reduction of weight gain and adiposity induced by CQ. For this, Wild-Type (WT) or UCP1-deficient mice fed a high fat diet will be treated with chloroquine or water and evaluated for weight gain, dietary intake, energy expenditure, adiposity, adipocyte diameter and number, insulin and glucose tolerance, glucose and fatty acid uptake and metabolism, mitochondrial content and function, insulin and inflammatory intracellular signaling, leukocyte recruitment and phenotype and inflammation of white adipose tissue and liver. We will also investigate during the doctorate the effect of the genetic inhibition of autophagy specifically on adipocytes in the metabolic, morphological and inflammatory alterations of adipose tissue due to Obesity. For this, wild-type mice (ULK1 fl/fl AdipoCre negative) or adipocyte-specific deletion of ULK1, a kinase that participates in autophagy initiation process (ULK1 fl/fl AdipoCre positive), will be induced to Obesity by HFD and evaluated for the parameters listed above. Appropriate statistical tests will be used to analyze the results (GraphPad Prism 6.0). Data will be expressed as mean ± SEM, level of significance p<0.05. (AU)