State Key Laboratory of Analytacal Chemistry for Life Science & Jiangsu Key Laboratory of Molecular Medicine, Medical school, Nanjing University, Nanjing, 210093, China.
Prenatal Diagnosis Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, 450052, China.
J Ovarian Res. 2018 Jan 10;11(1):6. doi: 10.1186/s13048-017-0375-7.
The polycystic ovary syndrome (PCOS) is a common metabolic and endocrine disorder with pathological mechanisms remain unclear. The following study investigates the ovarian hyperfibrosis forming via transforming growth factor-β (TGF-β) signaling pathway in Dehydroepiandrosterone (DHEA)- induced polycystic ovary syndrome (PCOS) rat model. We furthermore explored whether TGF-βRI inhibitor (SB431542) decreases ovarian fibrosis by counterbalancing the expression of fibrotic biomarkers.
Thirty female Sprague-Dawley rats were randomly divided into Blank group (n = 6), Oil group (n = 6), and Oil + DHEA-induced model group (n = 6 + 12). The model groups were established by subcutaneous injection of DHEA for 35 consecutive days. The 12 successful model rats were additionally divided in vehicle group (n = 6) and SB431542-treated group (n = 6). Vehicle group and SB431542-treated group, served as administration group and were intraperitoneally injected with DMSO and SB431542 for additional 14 consecutive days. Ovarian morphology, fibrin and collagen localization and expression in ovaries were detected using H&E staining, immunohistochemistry and Sirius red staining. The ovarian protein and RNA were examined using Western blot and RT-PCR.
In DHEA-induced ovary in rat, fibrin and collagen had significantly higher levels, while the main fibrosis markers (TGF-β, CTGF, fibronectin, a-SMA) were obviously upregulated. SB431542 significantly reduced the expression of pro-fibrotic molecules (TGF-β, Smad3, Smad2, a-SMA) and increased anti-fibrotic factor MMP2.
TGF-βRI inhibitor (SB431542) inhibits the downstream signaling molecules of TGF-β and upregulates MMP2, which in turn prevent collagen deposition. Moreover, ovarian hyperfibrosis in DHEA-induced PCOS rat model could be improved by TGF-βRI inhibitor (SB431542) restraining the transcription of accelerating fibrosis genes and modulating EMT mediator.
多囊卵巢综合征（PCOS）是一种常见的代谢和内分泌紊乱，其病理机制尚不清楚。本研究通过转化生长因子-β（TGF-β）信号通路探讨脱氢表雄酮（DHEA）诱导的多囊卵巢综合征（PCOS）大鼠模型中卵巢纤维化的形成。我们还探讨了 TGF-βRI 抑制剂（SB431542）是否通过平衡纤维化生物标志物的表达来减少卵巢纤维化。
30 只雌性 Sprague-Dawley 大鼠随机分为空白组（n=6）、油组（n=6）和油+DHEA 诱导模型组（n=6+12）。模型组连续皮下注射 DHEA 35 天。12 只成功建立模型的大鼠进一步分为溶剂组（n=6）和 SB431542 治疗组（n=6）。溶剂组和 SB431542 治疗组分别腹腔注射 DMSO 和 SB431542 连续 14 天。采用 H&E 染色、免疫组化和天狼猩红染色检测卵巢形态、纤维蛋白和胶原在卵巢中的定位和表达。采用 Western blot 和 RT-PCR 检测卵巢蛋白和 RNA。
在 DHEA 诱导的大鼠卵巢中，纤维蛋白和胶原水平明显升高，而主要的纤维化标志物（TGF-β、CTGF、纤维连接蛋白、α-SMA）明显上调。SB431542 明显降低了促纤维化分子（TGF-β、Smad3、Smad2、α-SMA）的表达，增加了抗纤维化因子 MMP2。
TGF-βRI 抑制剂（SB431542）抑制 TGF-β的下游信号分子，上调 MMP2，从而阻止胶原沉积。此外，TGF-βRI 抑制剂（SB431542）通过抑制加速纤维化基因的转录和调节 EMT 介质，可改善 DHEA 诱导的 PCOS 大鼠模型中的卵巢纤维化。
