GLP-1 Receptor Activation Inhibits Palmitate-Induced Apoptosis via Ceramide in Human Cardiac Progenitor Cells

J Clin Endocrinol Metab. 2017 Nov 1;102(11):4136-4147. doi: 10.1210/jc.2017-00970.

Abstract

Context: Increased apoptosis of cardiomyocytes and cardiac progenitor cells (CPCs) in response to saturated fatty acids (SFAs) can lead to myocardial damage and dysfunction. Ceramides mediate lipotoxicity-induced apoptosis. Glucagonlike peptide-1 receptor (GLP1R) agonists exert beneficial effects on cardiac cells in experimental models.

Objective: To investigate the protective effects of GLP1R activation on SFA-mediated apoptotic death of human CPCs.

Design: Human CPCs were isolated from cardiac appendages of nondiabetic donors and then exposed to palmitate with or without pretreatment with the GLP1R agonist exendin-4. Ceramide accumulation was evaluated by immunofluorescence. Expression of key enzymes in de novo ceramide biosynthesis was studied by quantitative reverse-transcription polymerase chain reaction and immunoblotting. Apoptosis was evaluated by measuring release of oligonucleosomes, caspase-3 cleavage, caspase activity, and terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling.

Results: Exposure of the CPCs to palmitate resulted in 2.3- and 1.9-fold higher expression of ceramide synthase 5 (CERS5) and ceramide desaturase-1, respectively (P < 0.05). This was associated with intracellular accumulation of ceramide and activation of c-Jun NH2-terminal protein kinase (JNK) signaling and apoptosis (P < 0.05). Both coincubation with fumonisin B1, a specific ceramide synthase inhibitor, and CERS5 knockdown prevented ceramide accumulation, JNK activation, and apoptosis in response to palmitate (P < 0.05). Exendin-4 also prevented the activation of the ceramide biosynthesis and JNK in response to palmitate, inhibiting apoptosis (P < 0.05).

Conclusions: Excess palmitate results in activation of ceramide biosynthesis, JNK signaling, and apoptosis in human CPCs. GLP1R activation counteracts this lipotoxic damage via inhibition of ceramide generation, and this may represent a cardioprotective mechanism.

MeSH terms

  • Apoptosis / drug effects*
  • Cells, Cultured
  • Ceramides / metabolism*
  • Exenatide
  • Glucagon-Like Peptide 1 / analogs & derivatives
  • Glucagon-Like Peptide-1 Receptor / agonists*
  • Humans
  • Myocytes, Cardiac / drug effects*
  • Myocytes, Cardiac / metabolism
  • Palmitic Acid / pharmacology*
  • Peptides / pharmacology*
  • Signal Transduction / drug effects
  • Spheroids, Cellular / drug effects
  • Spheroids, Cellular / metabolism
  • Stem Cells / drug effects*
  • Stem Cells / metabolism
  • Venoms / pharmacology*

Substances

  • Ceramides
  • Glucagon-Like Peptide-1 Receptor
  • Peptides
  • Venoms
  • Palmitic Acid
  • Glucagon-Like Peptide 1
  • Exenatide