Wang Fan, Zhang Zhenghong, Wang Zhaokai, Xiao Kaizhuan, Wang Qing, Su Jingqian, Wang Zhengchao
Provincial Key Laboratory for Developmental Biology and Neurosciences, College of Life Sciences (Qishan Campus), Fujian Normal University, 8, Shangsan Road, Fuzhou, 350007, China.
J Mol Histol. 2015 Apr;46(2):173-81. doi: 10.1007/s10735-015-9609-4. Epub 2015 Jan 23.
Polycystic ovary syndrome (PCOS) is a major health problem in reproductive-aged women worldwide, but the precise pathogenesis of PCOS remains unclear. Our previous study revealed that hypoxia-inducible factor (HIF)-1a mediated endothelin (ET)-2 signaling plays an important role in ovulation in rats. Therefore, the present study used a PCOS rat model to test the hypotheses that HIF-1a signaling is expressed and inhibited in ovaries during PCOS formation and that the HIF-1a/ET-2 signaling pathway is a target of dimethyldiguanide (DMBG) in the clinical treatment of PCOS. First, the development of a PCOS model and the effect of DMBG treatment were examined through ovarian histology and serum hormone levels, which were consistent with previous reports. Second, HIF-1a and ET-2 expression were detected by immunohistochemistry and western blot. The results showed decreased HIF-1a/ET-2 expression in the ovaries of PCOS rats, whereas DMBG treatment reversed the protein decreases and improved the PCOS symptoms. Third, to understand the molecular mechanism, HIF-1a/ET-2 mRNA expression was also examined. Interestingly, HIF-1a mRNA increased in the ovaries of PCOS rats, while ET-2 mRNA decreased, indicating that HIF-1a protein degradation may be involved in POCS development and treatment. Finally, HIF prolyl hydroxylase (PHD) activity was examined to further clarify the contribution of HIF-1a signaling to the development and treatment of PCOS. The results suggested that the inhibition of HIF-1a/ET-2 signaling may be caused by increased PHD activity in PCOS. DMBG-treated PCOS may further activate HIF-1a signaling at least partly through inhibiting PHD activity. Taken together, these results indicate that HIF-1a signaling is inhibited in a PCOS rat model through increasing PHD activity. DMBG treatment improved PCOS by rescuing this pathway, suggesting that HIF-1a signaling plays an important role in the development and treatment of PCOS. This HIF-1a-mediated ET-2 signaling pathway may be an important mechanism regulating PCOS formation and treatment in mammalian ovaries in vivo and should be a new clinical target for PCOS prevention and treatment in the future.
多囊卵巢综合征(PCOS)是全球育龄女性面临的一个主要健康问题,但PCOS的确切发病机制仍不清楚。我们之前的研究表明,缺氧诱导因子(HIF)-1α介导的内皮素(ET)-2信号传导在大鼠排卵过程中起重要作用。因此,本研究使用PCOS大鼠模型来验证以下假设:在PCOS形成过程中,HIF-1α信号在卵巢中表达并受到抑制,并且HIF-1α/ET-2信号通路是二甲双胍(DMBG)在PCOS临床治疗中的作用靶点。首先,通过卵巢组织学和血清激素水平检查PCOS模型的发育情况以及DMBG治疗的效果,结果与之前的报道一致。其次,通过免疫组织化学和蛋白质印迹法检测HIF-1α和ET-2的表达。结果显示,PCOS大鼠卵巢中HIF-1α/ET-2表达降低,而DMBG治疗逆转了蛋白质表达的降低并改善了PCOS症状。第三,为了解分子机制,还检测了HIF-1α/ET-2 mRNA的表达。有趣的是,PCOS大鼠卵巢中HIF-1α mRNA增加,而ET-2 mRNA减少,这表明HIF-1α蛋白降解可能参与了POCS的发生和治疗。最后,检测了HIF脯氨酰羟化酶(PHD)的活性,以进一步阐明HIF-1α信号传导对PCOS发生和治疗的作用。结果表明,PCOS中HIF-1α/ET-2信号的抑制可能是由于PHD活性增加所致。DMBG治疗的PCOS可能至少部分通过抑制PHD活性进一步激活HIF-1α信号。综上所述,这些结果表明,在PCOS大鼠模型中,HIF-1α信号通过增加PHD活性而受到抑制。DMBG治疗通过挽救该信号通路改善了PCOS,这表明HIF-1α信号在PCOS的发生和治疗中起重要作用。这种HIF-1α介导的ET-2信号通路可能是体内哺乳动物卵巢中调节PCOS形成和治疗的重要机制,并且应该成为未来PCOS预防和治疗的新临床靶点。