基于BDNF信号转导通路研究辅助生殖技术中卵巢储备功能降低的中药干预机制

卵巢储备功能降低（DOR）是影响辅助生殖技术(ART)成功率的主要瓶颈问题之一，目前西医学尚无公认有效的解决方案。本课题组一直致力于中药补肾调冲方防治DOR的临床及基础研究。本项目拟采用ELISA、TUNEL、免疫荧光共聚焦显微镜、定量PCR、Western Blot等技术，通过检测卵巢内各级卵泡数、血清INHB及AMH含量、细胞凋亡，卵母细胞线粒体及纺锤体等的分布、卵母细胞成熟相关因子表达、体外受精率及囊胚形成率等，研究补肾调冲方提高DOR小鼠卵巢储备功能，改善卵母细胞质量，提高受精率，促进未成熟卵母细胞体外成熟及胚胎发育能力的作用。在此基础上，进一步探讨脑源性神经生长因子（BDNF）信号转导通路异常与DOR发病的相关性，以及补肾调冲方通过干预BDNF信号通路治疗DOR的作用机制，为临床防治DOR以及提高ART的成功率提供新治疗方法，并为中医学"肾主生殖"等理论提供科学依据。

The occurrence of diminished ovarian reserve (DOR) has long been regarded a bottleneck restricting the success rate of ART (assisted reproductive technologies) procedures. So far western medicine has no real effective solution to this problem. Therefore, we propose the use of TCM as a complementary therapy to improve ovarian reserve. For years, our team has devoted itself to basic and clinical research concerning the effectiveness of Bushen Tiaochong Recipe on DOR prevention and treatment. In this project, we intend to explore the effects of Bushen Tiaochong Therapy on DOR mouse models. Using ELISA, TUNEL, quantitative PCR, Western Blot and immunofluorescence confocal microscopy, we counted the number of ovarian follicles at different developmental stages, measured serum INHB and AMH, detected apoptotic cells, investigated the distribution of oocyte mitochondria and spindle apparatus and observed the expression of oocyte maturation promoting factors, to see if the therapy can enhance oocytes quality, promote in vitro maturation of immature oocytes and repair the developmental ability of mouse embryos. Furthermore, the relevance between the abnormal BDNF (brain-derived neurotrophic factor) signaling pathway and the incidence of DOR was discussed and the mechanism of action of Bushen Tiaochong Therapy in treatment of DOR by altering the BDNF signaling pathway was explored. With this study we attempt to provide a new way for the clinical prevention and treatment of DOR and for the enhancement of ART success rate. Also, our work will provide theoretical supports for the TCM belief that "kidney governs reproduction" and "the theory of Chong and Ren".