EETs Reduction Contributes to Granulosa Cell Senescence and Endometriosis-Associated Infertility via the PI3K/AKT/mTOR Signaling Pathway

Adv Sci (Weinh). 2025 Nov;12(42):e05656. doi: 10.1002/advs.202505656. Epub 2025 Aug 19.

Abstract

We aimed to examine abnormal oxidative lipid levels and their related mechanisms in EM-associated infertility. Through liquid chromatography tandem mass spectrometry analysis, decreased levels of epoxyeicosatrienoic acids (EETs), which have antioxidant and anti-senescence effects are observed, in EM patient follicular fluid samples. EET levels are positively correlated with in vitro fertilization outcomes. Lower 14, 15-EET concentrations led to a decreased GC antioxidant capacity, reduced ATP production, reactive oxygen species (ROS) accumulation in oocytes, and abnormal cumulus-oocyte complex (COC) expansion, ultimately resulting in decreased fertility. Elevated soluble epoxide hydrolase (EPHX2) expression in EM-GCs is the main reason for EET reduction in EM follicular fluid. Inhibiting EPHX2 in vivo or in vitro can reverse these observed abnormalities by upregulating EETs. 14, 15-EET treatment alleviated GC senescence and improved fertility by inhibiting excessive PI3K/AKT/mTOR signaling pathway activation in EM-GCs, with BEZ-235-mediated inhibition of this pathway significantly alleviating ROS-induced cell senescence and abnormal COC expansion. Oxidative stress-induced decreased EZH2/H3K27Me3 histone methylation led to elevated EPHX2 expression patterns in EM-GCs. Decreased 14, 15-EET levels resulted in ROS accumulation, reduced EZH2 enzymatic activity, less EPHX2/H3K27Me3 histone methylation, and increased EPHX2 protein expression levels, which further reduced 14, 15-EET levels in a vicious feedback loop.

Keywords: 14, 15‐EET; EPHX2; EZH2/H3K27Me3; PI3K/AKT/mTOR signaling pathway; ROS; cellular senescence; endometriosis; ovary granulosa cell.

MeSH terms

  • 8,11,14-Eicosatrienoic Acid* / analogs & derivatives
  • 8,11,14-Eicosatrienoic Acid* / metabolism
  • Adult
  • Animals
  • Cellular Senescence* / physiology
  • Endometriosis* / complications
  • Endometriosis* / metabolism
  • Epoxide Hydrolases / metabolism
  • Female
  • Granulosa Cells* / metabolism
  • Humans
  • Infertility, Female* / etiology
  • Infertility, Female* / metabolism
  • Mice
  • Oxidative Stress
  • Phosphatidylinositol 3-Kinases / metabolism
  • Proto-Oncogene Proteins c-akt / metabolism
  • Reactive Oxygen Species / metabolism
  • Signal Transduction / physiology
  • TOR Serine-Threonine Kinases* / metabolism

Substances

  • Proto-Oncogene Proteins c-akt
  • TOR Serine-Threonine Kinases
  • Phosphatidylinositol 3-Kinases
  • Reactive Oxygen Species
  • MTOR protein, human
  • 8,11,14-Eicosatrienoic Acid
  • Epoxide Hydrolases