应用Gdf9新突变鼠模型进行卵源性卵巢低反应的机制研究

促排卵治疗已经有多年的历史，但仍然面临很多困境，如临床上棘手的预期和非预期卵巢低反应，而如何建立恰当的卵巢低反应模型是研究的瓶颈。本研究拟根据临床上新发现的GDF9基因突变类型，分别建立Gdf9基因无义突变和错义突变，以及双突变复合杂合小鼠模型，观察Gn依赖阶段中GDF9功能不同程度受损对卵泡颗粒细胞和卵泡膜细胞增殖、凋亡和甾体激素合成的影响，通过RNA测序和可溶性蛋白的液相芯片发现有差异的基因和卵泡旁分泌因子，并深入进行机制研究，探讨GDF9是否通过Smad通路与LH下游PKA/CREB/AP-1信号通路协同作用，从而影响抑制素A水平，最终干预卵泡的优势化过程。本研究将为阐明卵母细胞和卵泡体细胞之间的相互作用进一步提供理论支持，也为卵源性低反应的促排卵治疗提供新靶点和新策略。

Controlled ovarian stimulation, which has been applied for years, still faces many dilemmas, such as expected and unexpected poor ovarian response. How to establish an appropriate poor ovarian response model is the bottleneck for research in this field. Based on the newly discovered GDF9 gene mutations in our clinical practice, this study intends to establish the mouse models with homozygous nonsense mutation and homozygous missense mutation of GDF9 gene, as well as compound heterozygous mutation of Gdf9 gene. Using these models, we aim to investigate the impacts of different degrees of damages in GDF9 function on the proliferation, apoptosis and steroid hormone synthesis of Gn-dependent stage follicles. Changes in genes and paracrine factors inside the follicles will be studied by RNA sequencing and liquid chip. Moreover, this study will explore the mechanism of how GDF9 interacts with the downstream PKA/CREB/ AP-1 signaling pathway of LH through the Smad pathway, thereby affecting the level of Inhibin A in the follicle, and ultimately interfering with the dominant process of follicles. This study will provide further theoretical support to elucidate the interaction between oocytes and follicular cells, as well as new therapeutic targets and strategies for ovarian stimulation when facing oocyte-derived poor ovarian response.