Phospholipase C inhibits apoptosis of porcine primary granulosa cells cultured in vitro

J Ovarian Res. 2019 Sep 25;12(1):90. doi: 10.1186/s13048-019-0567-4.

Abstract

Phospholipase C (PLC) can participate in cell proliferation, differentiation and aging. However, whether it has a function in apoptosis in porcine primary granulosa cells is largely uncertain. The objective of this study was to examine the effects of PLC on apoptosis of porcine primary granulosa cells cultured in vitro. The mRNA expression of BAK, BAX and CASP3, were upregulated in the cells treated with U73122 (the PLC inhibitor). The abundance of BCL2 mRNA, was upregulated, while BAX and CASP3 mRNA expression was decreased after treatment with m-3M3FBS (the PLC activator). Both the early and late apoptosis rate were maximized with 0.5 μM U73122 for 4 h. The rate of early apoptosis was the highest at 4 h and the rate of late apoptosis was the highest at 12 h in the m-3M3FBS group. The protein abundance of PLCβ1, protein kinase C β (PKCβ), calmodulin-dependent protein kinaseII α (CAMKIIα) and calcineurinA (CalnA) were decreased by U73122, and CAMKIIα protein abundance was increased by m-3M3FBS. The mRNA expression of several downstream genes (CDC42, NFATc1, and NFκB) was upregulated by PLC. Our results demonstrated that apoptosis can be inhibited by altering PLC signaling in porcine primary granulosa cells cultured in vitro, and several calcium-sensitive targets and several downstream genes might take part in the processes.

Keywords: Apoptosis; Granulosa cells; Phospholipase C; Porcine.

MeSH terms

  • Animals
  • Apoptosis / drug effects*
  • Apoptosis / genetics
  • Calcineurin / genetics
  • Calcium / metabolism
  • Caspase 3 / genetics
  • Cell Proliferation / drug effects*
  • Cell Proliferation / genetics
  • Estrenes / pharmacology
  • Female
  • Gene Expression Regulation / drug effects
  • Granulosa Cells / drug effects
  • Granulosa Cells / metabolism*
  • Granulosa Cells / pathology
  • Phospholipase C beta / genetics
  • Phosphoprotein Phosphatases / genetics
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • Pyrrolidinones / pharmacology
  • Signal Transduction / drug effects
  • Sulfonamides / pharmacology
  • Swine
  • Type C Phospholipases / genetics*
  • bcl-2 Homologous Antagonist-Killer Protein / genetics
  • bcl-2-Associated X Protein / genetics

Substances

  • 2,4,6-trimethyl-N-(meta-3-trifluoromethylphenyl)benzenesulfonamide
  • Estrenes
  • Proto-Oncogene Proteins c-bcl-2
  • Pyrrolidinones
  • Sulfonamides
  • bcl-2 Homologous Antagonist-Killer Protein
  • bcl-2-Associated X Protein
  • 1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione
  • calmodulin dependent protein kinase II phosphatase
  • Calcineurin
  • Phosphoprotein Phosphatases
  • Type C Phospholipases
  • Phospholipase C beta
  • Caspase 3
  • Calcium