Priority Research Paper Volume 12, Issue 12 pp 11200—11223

Endothelin-1 induces cellular senescence and fibrosis in cultured myoblasts. A potential mechanism of aging-related sarcopenia

Elena Alcalde-Estévez1, *, , Ana Asenjo-Bueno2, *, , Patricia Sosa1, , Gemma Olmos1,3,4, , Patricia Plaza2, , María Ángeles Caballero-Mora5, , Diego Rodríguez-Puyol3,4,6, , María Piedad Ruíz-Torres1,3,4, *, , Susana López-Ongil2,3,4, *, ,

  • 1 Departamento Biología de Sistemas, Universidad de Alcalá, Alcalá de Henares, Madrid 28871, Spain
  • 2 Unidad de Investigación de la Fundación para la Investigación Biomédica del Hospital Universitario Príncipe de Asturias, Alcalá de Henares, Madrid 28805, Spain
  • 3 Instituto Reina Sofía de Investigación Nefrológica (IRSIN) de la Fundación Renal Iñigo Álvarez de Toledo (FRIAT), Madrid 28003, Spain
  • 4 Area 3-Fisiología y Fisiopatología Renal y Vascular del IRYCIS, Madrid 28046, Spain
  • 5 Servicio de Geriatría y Unidad de Fragilidad del Hospital de Ciudad Real, Ciudad Real 13003, Spain
  • 6 Servicio de Nefrología del Hospital Universitario Príncipe de Asturias, Alcalá de Henares, Madrid 28805, Spain
* Equal contribution

Received: December 11, 2019       Accepted: May 20, 2020       Published: June 22, 2020
How to Cite

Copyright © 2020 Alcalde-Estévez et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Endothelial dysfunction, with increased endothelin-1 (ET-1) synthesis, and sarcopenia, characterized by the loss of muscular mass and strength, are two aging–related conditions. However, a relationship between them has not been already established. The aim of this study was to determine whether ET-1 induces senescence and fibrosis in cultured murine myoblasts, which could be involved in the development of sarcopenia related to aging. For this purpose, myoblasts were incubated with ET-1 to assess cellular senescence, analyzed by senescence associated β-galactosidase activity and p16 expression; and fibrosis, assessed by fibronectin expression. ET-1 induced myoblast senescence and fibrosis through ETA receptor. The use of antioxidants and several antagonists revealed that ET-1 effect on senescence and fibrosis depended on ROS production and activation of PI3K-AKT-GSK pathway. To stress the in vivo relevance of these results, circulating ET-1, muscular strength, muscular fibrosis and p16 expression were measured in male C57Bl6 mice from 5-18-24-months-old. Old mice shown high levels of ET-1 correlated with muscular fibrosis, muscular p16 expression and loss of muscle strength. In conclusion, ET-1 promotes fibrosis and senescence in cultured myoblasts, similar results were found in old mice, suggesting a potential role for ET-1 in the development of sarcopenia related to aging.


AKT: tyrosine kinase B; C2C12: mouse myoblast cell line; ECE-1: endothelin-converting enzyme-1; ECM: extracellular matrix; ET-1: endothelin-1; FN: fibronectin; GSK: glycogen synthase kinase 3β; h: hour; ILK: integrin linked kinase; min: minute; NAC: N-acetylcysteine; PI3K: phosphoinositide 3 kinase; ROS: reactive oxygen species; SA-ß-GAL: senescence-associated ß-galactosidase.