TGFB1 modulates in vitro secretory activity and viability of equine luteal cells

Cytokine. 2018 Oct:110:316-327. doi: 10.1016/j.cyto.2018.03.038. Epub 2018 Apr 4.

Abstract

In the present report we describe the involvement of transforming growth factor B1 (TGF) in functional regression and structural luteolysis in the mare. Firstly, TGF and its receptors activin-like kinase (ALK) 5 and TGF receptor 2 were identified in corpus luteum (CL) steroidogenic, endothelial and fibroblast-like cells. Also, TGF and ALK5 protein expression were shown to be increased in Mid-, and Late-CL (p < 0.05). Subsequently, using an in vitro model with Mid-CL cells, we studied the role of TGF on secretory activity and cell viability. Cell treatment with TGF decreased progesterone (P4) and prostaglandin (PG) E2 concentrations in culture media (p < 0.05), and downregulated mRNA and protein of StAR, CYP11A1, cPGES and mPGES1 (p < 0.05). Conversely, TGF augmented PGF2a concentration in culture media, through PTGS2 and PGFS gene expression activation (p < 0.05). When cells were incubated with PGF2a, both TGF and ALK5 were upregulated (p < 0.05). Additionally, treatment with the pharmacological inhibitor of ALK5, ALK4 and ALK7 - SB431542 (SB) attenuated PGF2a functional and structural luteolytic actions. Indeed, SB blocked: (i) PGF2a inhibitory effect on StAR, CYP11A1, 3BHSD and mPGES1; (ii) PGF2a auto-amplification signal via PTGS2 and PGFS expression (p < 0.05); (iii) the PGF2a-induced BAX and FASL expression (p < 0.05). Finally, TGF decreased cell viability (p < 0.05) and promoted caspase 3 activity (p = 0.08) and the expression of pro-apoptotic FASL and BAX (p < 0.05). Our results suggest that TGF supports functional regression and structural luteolysis, and also confirm the importance of ALK5, ALK4 and ALK7 activation during PGF2a mediated luteolysis in mares.

Keywords: Apoptosis; Corpus luteum; Luteolysis; Steroidogenesis; Transforming growth factor B1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Activin Receptors, Type I / metabolism
  • Animals
  • Caspase 3 / metabolism
  • Cell Survival / physiology*
  • Cells, Cultured
  • Corpus Luteum / metabolism
  • Dinoprostone / metabolism
  • Down-Regulation / physiology
  • Female
  • Gene Expression / physiology
  • Horses
  • Luteal Cells / metabolism*
  • Luteolysis / metabolism
  • Progesterone / metabolism
  • RNA, Messenger / metabolism
  • Receptor, Transforming Growth Factor-beta Type I / metabolism
  • Transforming Growth Factor beta1 / metabolism*

Substances

  • RNA, Messenger
  • Transforming Growth Factor beta1
  • Progesterone
  • Activin Receptors, Type I
  • Receptor, Transforming Growth Factor-beta Type I
  • Caspase 3
  • Dinoprostone