Pigment epithelium derived factor PEDF exerts antioxidative effects in granulosa cells

Capsule:
Recombinant pigment epithelium–derived factor can protect granulosa cell viability in oxidative stress conditions by increasing SOD-1 and GPX- 2 mRNA levels, decreasing the protein level of BAX, and inducing AKT phosphorylation.

Authors:
Hadas Bar-Joseph, Ph.D., Ido Ben-Ami, M.D., Ph.D., Raphael Ron-El, M.D., Ruth Shalgi, Ph.D., Dana Chuderland, Ph.D.

Volume 102, Issue 3, Pages 891-898

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.

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