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  • Title: Pigment epithelium-derived factor exerts antioxidative effects in granulosa cells.
    Author: Bar-Joseph H, Ben-Ami I, Ron-El R, Shalgi R, Chuderland D.
    Journal: Fertil Steril; 2014 Sep; 102(3):891-898.e3. PubMed ID: 25064413.
    Abstract:
    OBJECTIVE: To determine whether supplementing granulosa cells cultures with pigment epithelium-derived factor (PEDF) can protect them from oxidative stress. DESIGN: We used either granulosa cell line or human primary granulosa cell culture from women undergoing in vitro fertilization (IVF) treatments. SETTING: University research facilities. ANIMAL(S): Imprinting control region female mice. INTERVENTION(S): 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. MAIN OUTCOME MEASURE(S): 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. RESULT(S): 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. CONCLUSION(S): This study demonstrates that PEDF represents a novel intrinsic antioxidant of granulosa cells.
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