In vitro study of the effects of endotelin-1 on irisine production by C2C12 cell derived myocytes

Abstract

Skeletal striated muscle is one of the most metabolically active tissues and therefore very susceptible to hormonal variations, be they endocrine, paracy and autocrine. This muscle is recognized as an endocrine organ that releases myocins, including irisine. This peptide acts as an energy-inducing signal, communicating directly with adipose tissue and triggering remodeling of subcutaneous adipocytes, modulating the metabolic profile and increasing energy expenditure throughout the body. Endothelin-1 (ET-1) is a pro-inflammatory and potent vasoconstrictor peptide that, when interacting with receptors on the vascular smooth muscle membrane, stimulates its contraction. However, the functional relationship between ET-1 and skeletal striated muscles is not yet clear. The main objective of this project is to study the effects of ET-1 on cell count, cell viability and irisin production in myocyte differentiated C2C12 cells. Therefore, 1000 cells / well (48 well plates) / 200 µL of culture medium will be cultured for 3, 5 and 7 days, in the absence and presence of two ET-1 concentrations (10-10 M and 10-6 M). At the end of each experimental time the cells will be counted (TC20" Bio-Rad counter), cell viability will be measured (MTT colorimetric assay) and irisine quantitation will be performed with an immunoenzyme assay (ELISA). Data will be analyzed by ANOVA and Fisher test. The significance level established will be 5% (p <0.05). It is hoped, with the results of this project, to understand in vitro interactions between ET-1 and irisine that may explain some physiological mechanisms of interest of our Laboratory that correlate endocrine-metabolic-reproductive signals.