Endothelin-1 induces lipolysis through activation of the GC/cGMP/Ca2+/ERK/CaMKIII pathway in 3T3-L1 adipocytes

Biochim Biophys Acta Mol Cell Biol Lipids. 2022 Feb;1867(2):159071. doi: 10.1016/j.bbalip.2021.159071. Epub 2021 Nov 5.

Abstract

Endothelin-1 (ET-1) is a potent vasoconstrictive peptide produced and secreted mainly by endothelial cells. Recent studies indicate that ET-1 can regulate lipid metabolism, which may increase the risk of insulin resistance. Our previous studies revealed that ET-1 induced lipolysis in adipocytes, but the underlying mechanisms were unclear. 3T3-L1 adipocytes were used to investigate the effect of ET-1 on lipolysis and the underlying mechanisms. Glycerol levels in the incubation medium and hormone-sensitive lipase (HSL) phosphorylation were used as indices for lipolysis. ET-1 significantly increased HSL phosphorylation and lipolysis, which were completely inhibited by ERK inhibitor (PD98059) and guanylyl cyclase (GC) inhibitor (LY83583). LY83583 reduced ET-1-induced ERK phosphorylation. A Ca2+-free medium and PLC inhibitor caused significant decreases in ET-1-induced lipolysis as well as ERK and HSL phosphorylation, and IP3 receptor activator (D-IP3) increased lipolysis. ET-1 increased cGMP production, which was not affected by depletion of extracellular Ca2+. On the other hand, LY83583 diminished the ET-1-induced Ca2+ influx. Transient receptor potential vanilloid-1 (TRPV-1) antagonist and shRNA partially inhibited ET-1-induced lipolysis. ET-1-induced lipolysis was completely suppressed by CaMKIII inhibitor (NH-125). These results indicate that ET-1 stimulates extracellular Ca2+ entry and activates the intracellular PLC/IP3/Ca2+ pathway through a cGMP-dependent pathway. The increased cytosolic Ca2+ that results from ET-1 treatment stimulates ERK and HSL phosphorylation, which subsequently induces lipolysis. ET-1 induces HSL phosphorylation and lipolysis via the GC/cGMP/Ca2+/ERK/CaMKIII signaling pathway in 3T3-L1 adipocytes.

Keywords: Calcium; Endothelin-1; Hormone-sensitive lipase; Lipolysis.

MeSH terms

  • 3T3-L1 Cells*
  • Adipocytes* / drug effects
  • Adipocytes* / metabolism
  • Animals
  • Calcium* / metabolism
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2 / metabolism
  • Cyclic GMP* / metabolism
  • Endothelin-1* / metabolism
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • Guanylate Cyclase / genetics
  • Guanylate Cyclase / metabolism
  • Lipolysis* / drug effects
  • Mice
  • Phosphorylation / drug effects
  • Signal Transduction / drug effects
  • Sterol Esterase / genetics
  • Sterol Esterase / metabolism

Substances

  • Endothelin-1
  • Cyclic GMP
  • Calcium
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2
  • Guanylate Cyclase
  • Sterol Esterase
  • Extracellular Signal-Regulated MAP Kinases