Cyclophosphamide (Cy), a chemotherapeutic agent, is widely used to treat tumoursand is also associated with premature ovarian insufficiency. 4-Hydroperoxycyclophosphamide (4-HC), an active metabolite of Cy, was used for in vitro experiments. Granulosa cells (GCs) are crucial for maintaining follicle development and are also used in reproductive toxicity research in vitro. Resveratrol (Res), a polyphenolic compound, exhibits multiple effects in cells and animal models. To date, whether Res pretreatment has a protective effect on GCs induced by Cy remains unclear. This was an in vitro study, and primary cultures of rat GCs were used. Rat GCs were treated with 4-HC alone, Res + 4-HC or Res + 4-HC  + EX527, and GCs survival rates, oxidative stress levels, apoptosis rates and related Sirt1 pathway proteins were evaluated. We demonstrated that 4-HC caused GC damage by increasing oxidative stress, autophagy and apoptosis. Res pretreatment improved 4-HC-induced GC damage by increasing Sirt1 expression, reducing oxidative stress levels and decreasing Beclin1, LC3B, Bax and Caspase-3 levels. Importantly, the addition of EX527, which is a selective inhibitor of Sirt1, reversed the protective effect of Res pretreatment, indicating that Sirt1 may be an important mediator of the protective effect of Res. Taken together, we demonstrated that Res may be a potential drug to improve fertility preservation for patients undergoing chemotherapy.

中文翻译：

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白藜芦醇预处理对环磷酰胺诱导的大鼠卵巢颗粒细胞损伤的保护作用：体外研究。

环磷酰胺（Cy）是一种化学治疗剂，被广泛用于治疗肿瘤，并且还与卵巢早衰有关。Cy的活性代谢物4-羟氧基环磷酰胺（4-HC）用于体外实验。颗粒细胞（GCs）对于维持卵泡发育至关重要，并且还用于体外生殖毒性研究。白藜芦醇（Res），一种多酚化合物，在细胞和动物模型中表现出多种作用。迄今为止，尚不清楚Res预处理是否对Cy诱导的GC具有保护作用。这是一项体外研究，使用了大鼠GC的原代培养物。大鼠GC仅用4-HC，Res + 4-HC或Res + 4-HC + EX527处理，并评估了GC的存活率，氧化应激水平，细胞凋亡率和相关的Sirt1途径蛋白。我们证明4-HC通过增加氧化应激，自噬和细胞凋亡引起GC损伤。Res预处理通过增加Sirt1表达，降低氧化应激水平并降低Beclin1，LC3B，Bax和Caspase-3水平来改善4-HC诱导的GC损伤。重要的是，添加EX527是Sirt1的选择性抑制剂，可以逆转Res预处理的保护作用，这表明Sirt1可能是Res保护作用的重要介体。两者合计，我们证明Res可能是提高接受化疗患者生育能力的潜在药物。重要的是，添加EX527是Sirt1的选择性抑制剂，逆转了Res预处理的保护作用，表明Sirt1可能是Res保护作用的重要介体。两者合计，我们证明Res可能是提高接受化疗的患者生育能力的潜在药物。重要的是，添加EX527是Sirt1的选择性抑制剂，逆转了Res预处理的保护作用，表明Sirt1可能是Res保护作用的重要介体。综上所述，我们证明Res可能是提高接受化疗的患者生育能力的潜在药物。