Rats with hyperandrogen-induced polycystic ovary syndrome (PCOS) have been shown to develop ovarian oxidative stress (OS) and fibrosis. The Sirt1 agonist, resveratrol, can reduce OS through inhibiting p66Shc in other models of OS. We created a rat PCOS model with increased OS levels following treatment with one of the two androgens, dehydroepiandrosterone (DHEA) and dihydrotestosterone (DHT). The PCOS related features were determined by measurement of malondialdehyde (MDA) and superoxide dismutase (SOD) levels or by examining the reactive oxygen species (ROS) levels using the DCF-DA probe. The potential mechanisms by which p66Shc/Sirt1 mediates ovarian fibrosis were explored by western blotting, quantitative reverse transcription-PCR, immunofluorescence staining, and immunohistochemistry. Hyperandrogen dramatically augmented OS and activation of fibrotic factors in the ovary. Our data demonstrated that treatment with resveratrol enhanced Sirt1 and decreased ovarian OS as well as inhibited phosphorylation of p66Shc both in vivo and in vitro. The treatment suppressed fibrotic factor activation and improved ovarian morphology. Lentivirus- or siRNA-mediated p66Shc knockdown resulted in a dramatic enhancement of Sirt1 expression, down-regulation of ROS and suppression of fibrotic factors in granulosa cells. Moreover, p66Shc overexpression markedly increased the expression of fibrotic factors. Additionally, silencing Sirt1 induced a dramatic increase in p66Shc and enhanced activation of fibrotic factors. p66Shc may be a direct target of Sirt1 for inducing ROS and thus promoting fibrosis. Further exploration of the mechanisms of p66Shc in both fibrosis and OS may provide novel therapeutic strategies that will facilitate the improvement in PCOS symptoms and reproductive functions.

中文翻译：

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p66Shc的抑制可防止雄激素过多引起的卵巢氧化应激和纤维化。

高雄激素性多囊卵巢综合征（PCOS）的大鼠已显示出卵巢氧化应激（OS）和纤维化。Sirt1激动剂白藜芦醇可以通过抑制其他OS模型中的p66Shc来降低OS。我们用两种雄激素之一，脱氢表雄酮（DHEA）和二氢睾酮（DHT）治疗后，创建了一个OS升高的大鼠PCOS模型。通过测量丙二醛（MDA）和超氧化物歧化酶（SOD）的水平或使用DCF-DA探针检查活性氧（ROS）的水平来确定PCOS的相关特征。p66Shc / Sirt1介导卵巢纤维化的潜在机制已通过蛋白质印迹，定量逆转录PCR，免疫荧光染色和免疫组织化学进行了探索。高雄激素可显着增强卵巢中的OS和纤维化因子的激活。我们的数据表明，在体内和体外，白藜芦醇治疗均可增强Sirt1并降低卵巢OS，并抑制p66Shc的磷酸化。该治疗抑制了纤维化因子激活并改善了卵巢形态。慢病毒或siRNA介导的p66Shc敲低可显着增强Sirt1表达，下调ROS并抑制颗粒细胞中的纤维化因子。此外，p66Shc过表达显着增加了纤维化因子的表达。此外，沉默Sirt1诱导p66Shc急剧增加并增强纤维化因子的激活。p66Shc可能是Sirt1的直接靶标，可诱导ROS从而促进纤维化。