Wang Daojuan, Wang Tingyu, Wang Rong, Zhang Xinlin, Wang Lei, Xiang Zou, Zhuang Lingjia, Shen Shanmei, Wang Hongwei, Gao Qian, Wang Yong
State Key Laboratory of Analytacal Chemistry for Life Science & Jiangsu Key Laboratory of Molecular Medicine, Medical School, Nanjing University, Nanjing, 210093, China.
Department of Cardiology, Affiliated Drum Tower Hospital, Nanjing University School of Medicine, 321 Zhongshan Road, 210008, Nanjing, Jiangsu Province, China.
J Transl Med. 2020 Feb 17;18(1):84. doi: 10.1186/s12967-020-02249-4.
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.
雄激素过多诱导的多囊卵巢综合征（PCOS）大鼠已被证明会出现卵巢氧化应激（OS）和纤维化。白藜芦醇作为Sirt1激动剂，在其他氧化应激模型中可通过抑制p66Shc来减轻氧化应激。
我们用脱氢表雄酮（DHEA）和双氢睾酮（DHT）这两种雄激素之一处理大鼠，创建了氧化应激水平升高的PCOS模型。通过测量丙二醛（MDA）和超氧化物歧化酶（SOD）水平，或使用DCF-DA探针检测活性氧（ROS）水平来确定PCOS相关特征。通过蛋白质免疫印迹法、定量逆转录PCR、免疫荧光染色和免疫组织化学探索p66Shc/Sirt1介导卵巢纤维化的潜在机制。
雄激素过多显著增强了卵巢中的氧化应激和纤维化因子的激活。我们的数据表明，白藜芦醇处理在体内和体外均增强了Sirt1并降低了卵巢氧化应激，同时抑制了p66Shc的磷酸化。该处理抑制了纤维化因子的激活并改善了卵巢形态。慢病毒或小干扰RNA介导的p66Shc基因敲低导致颗粒细胞中Sirt1表达显著增强、ROS下调以及纤维化因子受到抑制。此外，p66Shc过表达显著增加了纤维化因子的表达。另外，沉默Sirt1会导致p66Shc显著增加并增强纤维化因子的激活。
p66Shc可能是Sirt1诱导ROS从而促进纤维化的直接靶点。进一步探索p66Shc在纤维化和氧化应激中的机制可能会提供新的治疗策略，有助于改善PCOS症状和生殖功能。
