Abstract: The aim of this study was to investigate the function and mechanism of Oviductus Ranae (OR) on protecting rat ovarian granulosa cells from H₂O₂-induced oxidative damage.Granulosa cells were divided into six groups:control,model,positive control,as well as low,middle and high doses of OR-containing serum group.The granulosa cells were exposed to H₂O₂ for 2 h,and then treated with the OR-containing serum for 48 h.After that,the viability of granulosa cells was detected by CCK-8 assay.mRNA expressions of PI3K and AKT3 were assayed by real-time qPCR.The related proteins,such as NF-κB (p65),IκB-α, IKK-β,and p-Akt (Ser473),were detected by Western blotting.The results showed that the viability of granulosa cells was significantly decreased after stimulating H₂O₂. OR-containing serum can significantly increase the viability of granulosa cells.Furthermore,OR-containing serum can rescue the descending expression of PI3K and AKT3 mRNA induced by H₂O₂, and up-regulate the protein levels of p-Akt (Ser473),IKK-β and NF-κB (p65),and down-regulate the expression of IκBα.These results indicated that OR protected granulosa cells from oxidative stress-induced injury by regulating the PI3K/Akt/NF-κB signal pathway,which may be one of the antioxidant mechanism of OR.

Key words: Oviductus Ranae, ovarian granulosa cells, oxidative stress, H₂O₂