基于线粒体能量代谢研究补肾调肝法防治卵巢衰老的机制

In the elderly women with fertility needs, the decline in egg quality due to ovarian aging is the leading cause of infertility and assisted reproductive technology failure. Existing studies have shown that mitochondrial energy metabolism is the key to determine the quality of ootid.AMPK as a key element in the regulation of energy metabolism in the cell, the down-regulated expression of AMPK led to oocyte mitochondrial dysfunction, chromosomal dislocation. Basing on previous study of our team, we speculated that the decreased quality of ootid closely relate to the abnormalities of AMPK/ PGC-1α pathway in the process of ovarian aging.And it could been improved by the decoction of Wuyu. In this project, we focused on the function, tissue and molecular expression of ootid to observe the quality of , the number and structure of mitochondrial, spindle assembly, chromosome arrangement, intracellular ATP content in the young old mice body and in the immature ootid in vitro, respectively. We might not only reveal the mechanism which AMPK/PGC-1α affect the ootid quality by regulating the mitochondrial metabolism but also find the improving effect of the decoction of Wuyu. Finally, we will confirm the decoction of Wuyu is a kind of proconceptive drug in in elderly women.

有生育需求的高龄女性中，因卵巢衰老而出现的卵细胞质量下降是导致不孕及辅助生殖技术失败的主要原因。现有研究表明，线粒体能量代谢是决定卵细胞质量的关键。AMPK作为细胞内调节能量代谢的关键分子，其表达下调导致卵细胞线粒体功能异常、染色体错位。课题组推测：卵巢衰老过程中，卵细胞质量下降与AMPK/PGC-1α通路异常有关，补肾调肝中药乌萸汤可干预这一环节提高卵细胞质量。本项目利用初老小鼠及未成熟卵体外培养系统，通过研究卵细胞质量、线粒体数量及结构、纺锤体组装、染色体排列、细胞内ATP含量及相关分子表达的变化，从动物、组织、细胞、细胞器、分子等多层次系统揭示：在卵巢衰老过程中，AMPK/PGC-1α调控线粒体代谢影响卵细胞质量的机制及乌萸汤的干预作用，为乌萸汤在高龄女性助孕中的应用提供依据。

背景：随着个体年龄增长而出现的卵巢功能衰退，其实质是卵细胞数量减少、质量下降，常表现为月经周期紊乱，对卵巢刺激反应欠佳、高流产风险甚至不孕。AMPK能够调节卵细胞线粒体的能量代谢，其下游分子PGC-1α能通过调控NRFs的基因转录，调节线粒体氧化磷酸化功能。线粒体保持正常的数量和活性对于卵泡发育至关重要。.内容：本课题组体内实验以初老小鼠为研究对象，在中药乌萸汤干预后，分别从整体、组织、细胞、细胞器、分子生物学等不同层次分析乌萸汤对卵巢衰老的防治作用及机制；体外实验利用IVM技术，在未成熟卵体外培养过程中，结合AMPK激活剂AICAR、抑制剂Compound C作用下，明确AMPK/PGC-1α通路对线粒体功能及卵细胞质量的影响，明确乌萸汤在此过程中起到治疗作用的机制。.结果：随着小鼠年龄增大，卵母细胞AMPK/PCG-1α通路各基因及蛋白的表达受到影响，导致线粒体数目减少、膜电位变化减弱，产生的ATP减少、ROS增加，SOD酶活性降低，AMPKα、p-AMPKα、PGC-1α蛋白表达降低。乌萸汤可以通过参与AMPK/PCG-1α通路调控，增强线粒体能量代谢及氧化应激状态，从而促进卵细胞成熟，改善卵细胞质量。.意义：本课题组以线粒体能量代谢对卵细胞质量的影响为切入点，深入研究在卵巢衰老过程中，AMPK/PGC-1α通路调控线粒体能量代谢，影响卵细胞发育的具体机制，揭示乌萸汤对这一环节的干预作用，为乌萸汤及补肾调肝法在高龄女性助孕中的应用提供依据。