Critical Role of FoxO1 in Granulosa Cell Apoptosis Caused by Oxidative Stress and Protective Effects of Grape Seed Procyanidin B2

Figure 9

Knockdown of FoxO1 prevented GSPB2-induced autophagy. Cultured cells were transfected with FoxO1 shRNA or control scramble shRNA. Before being exposed to H₂O₂ (150 μM) for 6 h, the cells were pretreated with GSPB2 (10 μmol/L) or 3-MA (10 mmol/L) for 24 h. (a) Granulosa cell viability was tested after treatment. (b) Quantitative RT-PCR was employed to detect the relative expression of caspase-3. (c) Western blotting analysis of LC3-II protein in GSPB2-treated granulosa cells. (d) Quantification of the relative LC3-II protein level with densitometry. (e) Apoptosis was detected by a TUNEL assay. Scale bars are 100 μm. (f) Quantification of TUNEL-positive cells. Values are expressed as the mean ± SEM, ; , sh-Scr with GSPB2 versus sh-Scr control; versus sh-Scr with GSPB2 groups.

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