title: Pigment epithelium-derived factor exerts antioxidative effects in granulosa cells.
abstract: OBJECTIVE
  To determine whether supplementing granulosa cells cultures with pigment epithelium-derived factor (PEDF) can protect them from oxidative stress.
  
  
  METHODS
  We used either granulosa cell line or human primary granulosa cell culture from women undergoing in vitro fertilization (IVF) treatments.
  
  
  METHODS
  University research facilities.
  
  
  METHODS
  Imprinting control region female mice.
  
  
  METHODS
  Recombinant PEDF (rPEDF) was added to cultures of either primary granulosa cell culture or granulosa cell line in the present or absence of H2O2 triggering.
  
  
  METHODS
  We followed cell viability with the use of methylthiazolyl tetrazolium assay and tracked PEDF mechanism of action with the use of Western blot analysis, measuring the level of SOD-1 and GPX-1 mRNA, protein level of BAX, and phosphorylation of AKT.
  
  
  RESULTS
  We found that granulosa cell viability and the level of PEDF mRNA were both significantly reduced, in a dose-dependent manner, after exposure to H2O2. The rate of H2O2-induced apoptosis was significantly attenuated in granulosa cells treated with rPEDF. We showed that granulosa cells, of both humans and rodents, express the PEDF receptor, PNPLA2; once stimulated by rPEDF, the cells exhibited phosphorylation of AKT. Finally, we showed that PEDF exerts its antioxidative activity through the AKT signaling pathway.
  
  
  CONCLUSIONS
  This study demonstrates that PEDF represents a novel intrinsic antioxidant of granulosa cells.
authors: Department of Cell and Developmental Biology, Sackler Faculty of Medicine, Tel-Aviv University, Ramat-Aviv, Tel Aviv, Israel.
authors: Hadas
authors: H
authors: Bar-Joseph
authors: IVF and Infertility Unit, Department of Obstetrics and Gynecology, Assaf Harofeh Medical Center, Zerifin Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.
authors: Ido
authors: I
authors: Ben-Ami
authors: IVF and Infertility Unit, Department of Obstetrics and Gynecology, Assaf Harofeh Medical Center, Zerifin Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.
authors: Raphael
authors: R
authors: Ron-El
authors: Department of Cell and Developmental Biology, Sackler Faculty of Medicine, Tel-Aviv University, Ramat-Aviv, Tel Aviv, Israel.
authors: Ruth
authors: R
authors: Shalgi
authors: Department of Cell and Developmental Biology, Sackler Faculty of Medicine, Tel-Aviv University, Ramat-Aviv, Tel Aviv, Israel. Electronic address: chuderla@gmail.com.
authors: Dana
authors: D
authors: Chuderland
mesh_terms: D000328:Adult; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D005260:Female; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D051379:Mice; D009414:Nerve Growth Factors / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D018384:Oxidative Stress; D051381:Rats; D015843:Serpins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D055815:Young Adult; D000099174:Pigment Epithelium-Derived Factor
chemical_list: D005136:Eye Proteins; D006861:Hydrogen Peroxide; D009414:Nerve Growth Factors; D015843:Serpins; D000099174:Pigment Epithelium-Derived Factor
keywords: AKT; H(2)O(2); PEDF; granulosa; oxidative stress