Periovulatory follicular fluid levels of estradiol trigger inflammatory and DNA damage responses in oviduct epithelial cells

PLoS One. 2017 Feb 23;12(2):e0172192. doi: 10.1371/journal.pone.0172192. eCollection 2017.

Abstract

Objective: Ovarian steroid hormones (mainly E2 and P4) regulate oviduct physiology. Serum-E2 acts on the oviduct epithelium from the basolateral cell compartment. Upon ovulation, the apical compartment of the oviduct epithelium is temporarily exposed to follicular fluid, which contains much higher levels of E2 than serum. The aim of this study was to evaluate the effects of human periovulatory follicular fluid levels of E2 on oviduct epithelial cells using two porcine in vitro models.

Methods: A cell line derived from the porcine oviductal epithelium (CCLV-RIE270) was characterized (lineage markers, proliferation characteristics and transformation status). Primary porcine oviduct epithelial cells (POEC) were cultured in air-liquid interface and differentiation was assessed histologically. Both cultures were exposed to E2 (10 ng/ml and 200 ng/ml). Proliferation of CCLV-RIE270 and POEC was determined by real-time impedance monitoring and immunohistochemical detection of Ki67. Furthermore, marker gene expression for DNA damage response (DDR) and inflammation was quantified.

Results: CCLV-RIE270 was not transformed and exhibited properties of secretory oviduct epithelial cells. Periovulatory follicular fluid levels of E2 (200 ng/ml) upregulated the expression of inflammatory genes in CCLV-RIE270 but not in POEC (except for IL8). Expression of DDR genes was elevated in both models. A significant increase in cell proliferation could not be detected in response to E2.

Conclusions: CCLV-RIE270 and POEC are complementary models to evaluate the consequences of oviduct exposure to follicular fluid components. Single administration of periovulatory follicular fluid E2 levels trigger inflammatory and DNA damage responses, but not proliferation in oviduct epithelial cells.

MeSH terms

  • Animals
  • Cell Line
  • Cell Proliferation
  • Cells, Cultured
  • DNA Damage*
  • DNA Repair
  • Epithelial Cells / cytology
  • Epithelial Cells / immunology*
  • Epithelial Cells / metabolism
  • Estradiol / immunology*
  • Female
  • Follicular Fluid / immunology*
  • Follicular Fluid / metabolism
  • Gene Expression Regulation
  • Inflammation / genetics*
  • Inflammation / immunology
  • Oviducts / cytology*
  • Oviducts / immunology
  • Oviducts / metabolism
  • Swine

Substances

  • Estradiol

Grants and funding

SEPV was supported by Dahlem Research School (FU Berlin), Germany. SC was supported by the Nature Science Funds from Hebei Ministry of Education (QN2014124), China.