1. Academic Validation
  2. Suppression of p66Shc prevents hyperandrogenism-induced ovarian oxidative stress and fibrosis

Suppression of p66Shc prevents hyperandrogenism-induced ovarian oxidative stress and fibrosis

  • J Transl Med. 2020 Feb 17;18(1):84. doi: 10.1186/s12967-020-02249-4.
Daojuan Wang 1 Tingyu Wang 1 Rong Wang 1 Xinlin Zhang 2 Lei Wang 1 Zou Xiang 3 Lingjia Zhuang 1 Shanmei Shen 4 Hongwei Wang 1 Qian Gao 5 Yong Wang 6
Affiliations

Affiliations

  • 1 State Key Laboratory of Analytacal Chemistry for Life Science & Jiangsu Key Laboratory of Molecular Medicine, Medical School, Nanjing University, Nanjing, 210093, China.
  • 2 Department of Cardiology, Affiliated Drum Tower Hospital, Nanjing University School of Medicine, 321 Zhongshan Road, 210008, Nanjing, Jiangsu Province, China.
  • 3 Department of Health Technology and Informatics, Faculty of Health and Social Sciences, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong, China.
  • 4 Department of Endocrinology, The Affiliated Drum Tower Hospital, Medical School, Nanjing University, Nanjing, 210093, China.
  • 5 State Key Laboratory of Analytacal Chemistry for Life Science & Jiangsu Key Laboratory of Molecular Medicine, Medical School, Nanjing University, Nanjing, 210093, China. qian_gao@nju.edu.cn.
  • 6 State Key Laboratory of Analytacal Chemistry for Life Science & Jiangsu Key Laboratory of Molecular Medicine, Medical School, Nanjing University, Nanjing, 210093, China. yongwang@nju.edu.cn.
Abstract

Background: 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.

Methods: 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.

Results: 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.

Conclusions: 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.

Keywords

Fibrotic factors; Granulosa cells; PCOS; Reactive oxygen species; p66Shc.

Figures
Products