Ovarian insufficiency such as premature ovarian failure and early menopause is an emerging problem in the treatment of female reproduction. The development of alternative therapeutic agents is essential to restore ovarian function. Resveratrol is a polyphenol complex known to have different effects in various tissues and cells under physiological and pathological conditions. In cancer cells, resveratrol has potent anti-inflammatory and antioxidant effects and has been found to induce apoptosis and cell cycle arrest. Bezerra et al. found that RES promotes primordial follicle activation by reducing DNA breaks and stimulates granulosa cell proliferation by activating the PI3K pathway. In the arat model of POI, Li et al. RES inhibited granulosa cell proliferation by activating the PI3K/AKT signaling pathway to inhibit granulosa cell apoptosis.
To investigate the effect of resveratrol against oxidative stress in POF ovaries, we measured the mRNA levels or activities of key antioxidant enzymes in POF ovaries and RES group ovaries. The results showed that the expression levels of SOD2 and CAT were decreased in the POF group compared with the control group, while the expression levels of GPx, SOD2, and CAT were significantly upregulated in the 40 mg/kg RES group. the total SOD and SOD2 enzyme activities were significantly lower in the POF group than in the control group, and the activity levels of the other two key antioxidant enzymes, GPx and CAT, were also significantly lower in the POF group than in the control group. Total SOD, SOD2, GPx, and CAT levels were significantly higher in the RES-treated group than in the POF group. In addition, the levels of MDA and ROS in the POF group were significantly upregulated and gradually decreased after RES treatment. Further, we isolated POF ovaries and incubated them with 1um RES in vitro for 72 h. Corresponding to the in vivo changes, the mRNA expression levels of the three antioxidant enzymes were significantly decreased in the POF group compared with the normal group, and RES treatment significantly restored the enzyme activities in POF ovaries. RT-PCR and Western blot analysis showed that compared with POF ovaries, RES treatment partially but significantly increased the expression levels of Moh and Oct4, suggesting a rescue effect of RES on germ-line cells in the POF model.
To optimize the concentration, we selected five different doses of RES co-cultured with fgcs at 0.1, 0.5, 1.5, and 10 μm in the CCK-8 method. compared with the normal group, the activity of FGSCs was slightly enhanced in the 0.1 μm and 5 μm groups, significantly increased in the 0.5 μm and 1 μm groups, and decreased in the 10 μm group, indicating that 0.5 μm and 1 RES were suitable concentrations for FGSC proliferation. To investigate the protective effect of resveratrol on FGCS, RES was co-cultured with FGCS pretreated with 40 bu for 48 h to simulate the POF model. After 72 h of RES treatment, the SOD2 level in the 0.5 group was slightly higher than that in the BU group, while that in the 1 um group was significantly higher than that in the BU group. the MDA concentration in the RES group was significantly lower than that in the BU group and higher than that in the normal group. Although BU significantly inhibited the proliferation of FGSC, RES effectively increased the activity of fgcs in CCK-8 analysis. Western blotting experiments showed that Mvh and Oct4 expression levels were significantly downregulated in the BU group, and this change was reversed after 72h of RES treatment.