Effect of early outgrowth cells in apoptosis of cultured podocytes exposed to high glucose or hydrogen peroxide

• Grant number: 13/05156-9
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Start date: September 01, 2013
• End date: February 28, 2014
• Field of knowledge: Biological Sciences - Physiology
• Principal Investigator: Jose Butori Lopes de Faria
• Grantee: Elisa Mauro Peixoto Prado
• Supervisor: Richard E. Gilbert
• Host Institution: Faculdade de Ciências Médicas (FCM). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Institution abroad: University of Toronto (U of T), Canada
• Associated to the scholarship: 10/05841-5 - Oxidative stress contribution for glomerular filtration barrier integrity in diabetic and hipertensive rats, BP.PD

Abstract

Oxidative stress induced by hyperglycaemia causes alteration in the glomerular filtration barrier, thereby leading to albuminuria, which is characteristic of diabetic nephropathy (DN). Podocytes are an important component of the integrity of the glomerular filtration barrier. Treatment with antioxidants diminishes diabetes-induced podocyte apoptosis and is therefore a potential therapeutic tool for DN. Early outgrowth cells (EOCs) attenuate reactive oxygen species (ROS) production and reduce albuminuria in db/db mice. The aim of this study is to investigate if EOCs are able to improve podocyte function, particularly due to podocyte apoptosis induced by high glucose or oxidative stress. We will grow a human podocyte cell line under normal glycaemia and hyperglycaemia, oxidative stress condition (H2O2 treatment), in the presence or absence of EOCs's -conditioned medium, siRNA for Nrf2 protein, and inhibitor of SIRT-1 pathway. We also co-culture EOCs with podocytes. Analysis of podocyte apoptosis will be done by fluorometric terminal deoxynucleotidyl transferase-mediated nick-end labelling (TUNEL) and caspase-3 activity, and podocyte adhesion will be estimated. Understanding the mechanism of action of EOCs in proteinuria of DN constitutes the basis for using this technique to treat DN. (AU)