To investigate the effect of Yishen Shugan Decoction(YSD)-containing serum on apoptosis of granulosa cells in rats treated with Tripterygium wilfordii glycosides(TWG) and the underlying mechanism.Methods:Ovarian granulosa cells of rats were isolated and treated with different concentration of YSD-containing serum and different concentration of TWG-containing serum for 48 h.Cell survival rate was detected by Cell Counting Kit-8(CCK-8) assay，and the doses of YSD-containing serum and TWG-containing serum were screened.Cells were classified into normal group(normally cultured cells)，injury group，low-dose，medium-dose，and high-dose Chinese medicine groups，and positive control group(estradiol-containing serum).Cell proliferation，apoptosis，and expression of phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR) pathway-related proteins were detected by CCK-8 assay，terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL) staining，and Western blotting.Results:Cell proliferation，and expression of B-cell lymphoma-2(Bcl-2)，phosphorylated(p)-PI3K，p-AKT，and p-mTOR were inhibited(P<0.05)，and the apoptosis rate and expression of Bcl-2-associated X protein(Bax) and cleaved(cl)-caspase-3 were increased(P<0.05) in the injury group compared with those in the normal group.Cell proliferation，and expression of Bcl-2，p-PI3K，p-AKT，and p-mTOR were increased(P<0.05) and apoptosis rate，and expression of Bax and cl-caspase-3 were inhibited(P<0.05) in positive control group，and low-dose，medium-dose，and high-dose Chinese medicine groups as compared with those in the injury group.The above increase and inhibition were in a dose-dependent manner in the Chinese medicine groups.Conclusion:YSD-containing serum may inhibit the apoptosis of ovarian granulosa cells by activating PI3K/AKT/mTOR pathway.