补肾法改善重复COS卵母细胞线粒体功能与调控Nrf2抗氧化应激通路的关系

Traditional Chinese medicine (TCM) considers that kidney store essence and dominate reproduction. The TCM treatment of “kidney-invigorating” focuses on improving the reproductive function. Oocyte is the essence of reproduction and essential for embryonic development. The mitochondria are known as the powerhouse of the cell, and their dysfunction is associated with oxidative stress. Our studies that supported by previous two funds (30772818，81173294) confirmed that kidney-invigorating treatment could increase the quality of oocyte and improve the oxidative stress of ovary. Pre-experimental results indicated that kidney-invigorating treatment could improve oxidative stress of ovary and mitochondrial function of oocyte in mice with embryo implantation dysfunction induced by repeated control ovarian stimulation (COS). Therefore, we speculate that kidney-invigorating treatment may increase antioxidant ability of oocyte and its microenvironment, then improve mitochondrial function, thus to enhance the ability of oocyte. The project intends to observe the effects of repeated COS on the oxidative stress of follicular fluid and granulosa cells, and further to investigate whether kidney-invigorating treatment could improve the oxidative stress at clinical level. In addition, we will establish an animal model with decreasing the quality of oocyte by repeated COS and study the effects of kidney-invigorating treatment on the oxidative stress and mitochondrial function of oocyte. The transcription factor nuclear factor erythroid 2 related factor 2 (Nrf2) is the master regulator of the cellular redox homeostasis which is associated with mitochondrial function. So we will further study the mechanism of kidney-invigorating treatment improving mitochondrial function via regulating Nrf2 antioxidant stress pathway at the cellular level. This study have significant implications for exploring the mechanism of traditional Chinese medicine in improving the quality of oocyte and illuminating the theory of “kidney dominating reproduction”.

肾藏精，主生殖，卵母细胞乃生殖之精，是胚胎发育的关键。线粒体变化是评估卵母细胞质量的重要标志，氧化应激是影响卵母细胞线粒体功能的核心因素。前两项基金（30772818，81173294）研究证实，补肾法具有提高卵母细胞质量和改善卵巢氧化损伤的作用。预实验结果显示，补肾法能够改善重复COS小鼠卵巢氧化应激状态和卵母细胞线粒体功能。因此我们推测，补肾法通过提高卵母细胞及周围微环境抗氧化能力，进而改善卵母细胞线粒体功能，发挥提高卵母细胞质量的作用。本项目拟通过临床研究，研究重复COS对卵母细胞周围微环境卵泡液及颗粒细胞氧化应激的影响和补肾法对其氧化应激的改善作用；建立重复COS小鼠模型，研究补肾法对模型小鼠卵巢、卵母细胞氧化应激及卵母细胞线粒体功能的影响；建立氧化应激细胞模型，研究补肾法对Nrf2抗氧化应激通路的调控作用。以期阐明补肾法提高卵母细胞质量的分子机制，丰富中医“肾主生殖”的科学内涵。

不孕症发病率的逐年升高，体外受精-胚胎移植（in vitro fertilization and embryo transfer, IVF-ET）技术是目前治疗不孕症的重要手段，控制性卵巢刺激（control ovarian stimulation, COS）是IVF过程中关键步骤。受妊娠率不理想的限制，患者往往要进行反复COS，而这会造成卵巢功能、卵母细胞质量下降。本项目根据“肾主生殖”理论，进行补肾法改善重复COS患者卵母细胞质量的研究。临床研究证实，补肾法可改善临床重复COS患者卵泡液及卵巢颗粒细胞的氧化应激及颗粒细胞线粒体功能，进而改善患者优质胚胎率及临床妊娠结局；动物实验证实，补肾法可改善重复COS模型小鼠卵巢和卵母细胞氧化应激，提高卵母细胞线粒体功能；细胞研究证实，补肾法通过调控PKC/Nrf2、PI3K-Akt/Nrf2信号通路发挥抗氧化作用，改善卵母细胞线粒体功能，进而提高卵母细胞质量、成活率及第一极体排出率。这对于阐释中医调经种子作用机制、丰富“肾主生殖”理论内涵、完善中医生殖理论具有重要的意义。