补肾法改善重复卵巢刺激卵母细胞线粒体功能与调控Sirt3信号通路的关系

Traditional Chinese medicine (TCM) considers that kidney store essence and dominate reproduction. Kidney invigorating is a common treatment for infertility. Mitochondria are the marker to evaluate the quality of oocyte, and oxidative stress has become an important factor affecting oocyte. And oxidative stress has become an important factor affecting oocyte. With the increasing incidence of infertility, IVF-ET has become the main way to treat infertility. Ovarian stimulation (OS) is an important step in IVF-ET. Repeated OS is very common in clinical. However, repeated OS can cause decrease of pregnancy rate, because of decrease of oocyte quality. The oxidative stress state of oocytes was confirmed after repeated OS in Foundation (8177151342). Sirtuin3 (Sirt3) belongs to Sirtuin, the main protein deacetylase in the mitochondria. Could Bushen Tiaojing Decoction improve the quality of oocytes by Sirt3-LKB1-AMPK signal pathway? It is worth studying. In animal experiments we observe the ultrastructural changes of oocytes by repeated OS. And on the basis of this, to investigate the role of Sirt3-LKB1-AMPK signaling pathway in the improvement of oocyte oxidative stress. At the cell experiments, through overexpression and knockout Sirt3, to study the specific mechanism of Bushen Tiaojing Decoction to improve the quality of oocyte. This is of great significance for enriching the theoretical connotation of " kidney dominate reproduction ".

肾藏精，主生殖，卵母细胞质量受肾精影响，补肾法是临床治疗不孕症的常用治法。线粒体是评估卵母细胞质量的标志，氧化应激成为影响卵母细胞的重要因素。在研基金（8177151342）结果已证实重复卵巢刺激后存在卵母细胞氧化应激状态。Sirtuin3（Sirt3）属于Sirtuin是线粒体内主要的蛋白去乙酰化酶，LKB1调节细胞能量代谢，AMPK能够调节体内能量平衡，补肾调经方能否通过调控Sirt3-LKB1-AMPK信号通路提高卵母细胞质量？值得进一步研究。本研究拟在动物实验中观察重复卵巢刺激对卵母细胞超微结构变化，并以此为基础，探讨补肾法诱导Sirt3-LKB1-AMPK信号通路在改善卵母细胞氧化应激中的作用。在细胞水平，通过过表达和敲低Sirt3，从正反两方面验证Sirt3在改善卵母细胞氧化应激，探讨改善卵母细胞质量的具体机制。这对于丰富“肾主生殖”的理论内涵具有重要意义。

肾藏精，主生殖，卵母细胞质量受肾精影响，补肾法是临床治疗不孕症的常用治法。前期研究结果证实，以补肾调经方为代表方的补肾法能够提高重复卵巢刺激后的卵母细胞质量，改善妊娠结局。线粒体是为卵母细胞及早期胚胎供能的重要细胞器。预实验结果也证实，补肾调经方可改善重复卵巢刺激后的卵母细胞线粒体功能，是提高卵母细胞质量的机制之一。Sirtuin3（Sirt3）属于Sirtuin家族，是线粒体内主要的蛋白去乙酰化酶，LKB1调节细胞能量代谢，AMPK能够调节体内能量平衡。由此我们提出假设，补肾法可能通过调控Sirt3-LKB1-AMPK信号通路改善氧化应激提高卵母细胞线粒体功能。本项目通过在体实验验证，补肾法可改善重复卵巢刺激造成的卵母细胞超微结构异常（包括线粒体功能、纺锤体/染色体、微丝、皮质颗粒），提高卵母细胞质量。细胞实验中，我们对小鼠卵母细胞进行体外培养，通过过表达及抑制Sirt3的表达，探讨了补肾法调控Sirt3-LKB1-AMPK信号通路，改善抗氧化酶MnSOD的表达，提高卵母细胞线粒体功能的作用机制。这对于揭示中医“肾主生殖”的理论内涵，探索中医调经种子的作用机制具有重要意义。