补肾健脾法基于SCFA-FOXO1途径调控卵巢颗粒细胞自噬机制及其改善PCOS子代发育作用研究

补肾健脾法是中医学治疗多囊卵巢综合征（PCOS）不孕的重要治法，申请人前期发现该治法能显著改善高雄性PCOS不孕患者的妊娠结局，降低妊娠期合并症发生，但其机制有待深入研究。补肾健脾法能显著回调PCOS模型大鼠卵巢颗粒细胞FOXO1等自噬相关基因的异常表达，提示其可能调控颗粒细胞自噬而影响卵泡与卵母细胞发育，从而为揭示中医治疗PCOS机制提供了新思路。本项目拟综合运用双荧光活细胞成像、ChIP等技术研究补肾健脾法对SCFA-FOXO1途径的调控机制，从影响卵巢颗粒细胞自噬行为的角度，探讨其改善高雄性PCOS不孕患者促排卵妊娠结局的相关分子机制；在此基础上，通过靶向代谢组学、胚胎实时监测等技术探讨补肾健脾法对PCOS大鼠卵母细胞、早期胚胎和子代发育的影响与分子机制。这不仅对于揭示中医学“天癸”学说和“脾肾同治”理论的科学基础具有重要意义，对发展中医干预PCOS的治疗策略也具有显著的临床价值。

Bu-Shen-Jian-Pi is an important traditional Chinese medicine therapeutic principle in treating infertility of polycystic ovary syndrome (PCOS). The applicant found that Bu-Shen-Jian-Pi can significantly improve the pregnancy outcomes and reduce the incidence of pregnancy complications of hyperandrogenic PCOS women, however, the underlying mechanism needs further study. Bu-Shen-Jian-Pi can positively regulate the abnormal expression levels of important genes associated with autophagy, such as forkhead box O1 (FOXO1), in the ovarian granulosa cells of PCOS model rats, indicating it may be through regulating autophagy in the granulosa cells that Bu-Shen-Jian-Pi plays important roles in the development of follicles and oocytes, which provides a new way to reveal the mechanism of traditional Chinese medicine in treating PCOS. The present project will explore whether it is through regulating autophagy in ovarian granulosa cells mediated by SCFA-FOXO1 pathway that Bu-Shen-Jian-Pi improves the pregnancy outcomes by comprehensively using transmission electron microscopy, double fluorescence living cell imaging, etc. Based above, the project will also investigate the effects and underlying mechanism of Bu-Shen-Jian-Pi on the development of oocytes, early embryos and offspring of hyperandrogenic PCOS model rats using targeted metabonomics technology, embryo real-time monitoring system,etc. The present study has great significance to reveal the scientific basis of theories of "Tian Gui" and "Pi Shen Tong Zhi", and has eminent clinical significance for developing the protocol of traditional Chinese medicine in treating PCOS.