逍遥散对慢性应激肝郁证卵泡发育障碍的作用和卵巢局部BDNF调控机制研究

Follicular maldevelopment induced by chronic stress is associated with various common reproductive endocrine disorders. Liver depression is the most common traditional Chinese medicine syndrome induced by chronic stress. Xiaoyao powder is a famous gynecological prescription to treat this syndrome. Numerous reseaches showed that xiaoyao powder could protect multiple systems and organs in animal model of liver depression syndrome induced by chronic stress. However, little is known about the machnisms and the effects of Xiaoyao powder on the follicular maldevelopment induced by chronic stress. On the basis of literature review, our preliminary clinic studies and the researches supported by national natural science fundation of China, we proposed that Xiaoyao powder may ameliorate follicular maldevelopment induced by chronic stress by up-regulation of ovarian BDNF. To test our hypotheses, a classic stress model induced by chronic unpredictable mild stress is going to be used. We will observe the effect of Xiaoyao powder on behavior, ovulation, ovarian endocrine, oocytes developmental potential, general morphology, ultrastructure, follicular cell proliferation and apoptosis, as well as the level and distribution of BDNF. BDNF antibody and inhibitor of TrkB receptor were used to validate the role and pathway of BDNF. The study would help to elucidate the ovarian BDNF mechanisms and the effects of xiaoyao powder on follicular maldevelopments with liver depression syndrome induced by chronic stress. The study is intended to provide a theory basis of modern molecular biology and a new idea in the traditional Chinese medicine therapy of stress-associated psychosomatic diseases.

慢性应激可致卵泡发育障碍，与多种妇科常见生殖和内分泌紊乱疾病密切相关。女性慢性应激中医证候多见肝郁证，妇科名方逍遥散对女性肝郁证疗效显著。研究显示该方可对慢性应激肝郁证模型动物多系统多器官起保护作用，但目前尚无逍遥散对慢性应激所致卵泡发育障碍的作用及机制报道。在综合文献、前期临床研究及国家青年基金研究基础上，我们提出逍遥散可通过卵巢局部BDNF调控因子改善慢性应激肝郁证卵泡发育障碍。本研究将采用慢性不可预见应激肝郁证小鼠模型，观察逍遥散对慢性应激小鼠行为学、排卵、卵泡内分泌、卵母细胞发育潜能、一般形态学、超微结构、卵泡细胞增殖和凋亡以及BDNF表达和分布的影响，并运用BDNF抗体和TrkB受体抑制剂等工具药验证BDNF的作用和信号途径。研究旨在阐明逍遥散对慢性应激肝郁证卵泡发育障碍的作用及其卵巢局部BDNF调控机制，为慢性应激相关女性心身疾病的中医药治疗提供现代分子生物学理论基础和新思路。

背景：慢性应激可致卵泡发育障碍，与多种常见女性生殖内分泌紊乱疾病相关。慢性应激中医证候多为肝郁证，针对该证的妇科名方逍遥散疗效显著。研究显示该方可对慢性应激肝郁证动物多系统多器官起保护作用，但目前尚无逍遥散对慢性应激所致卵泡发育障碍的作用及机制报道。在综合文献、前期临床研究及国家自然科学基金研究基础上，我们提出逍遥散可通过上调卵巢局部调控因子BDNF改善慢性应激肝郁证卵泡发育障碍。主要研究内容：本研究将采用慢性不可预见温和应激小鼠模型，观察逍遥散对慢性应激小鼠行为学、排卵、卵巢内分泌、卵母细胞发育潜能、一般形态学、超微结构、卵泡细胞增殖和凋亡以及BDNF表达的影响，并运用BDNF抗体反向验证BDNF的作用。重要结果和数据：我们的研究发现，慢性应激可致小鼠排卵数减少、分泌雌二醇和孕酮水平下降、卵泡发育异常、卵母细胞超微结构异常，使颗粒细胞增殖减少，凋亡增加，使BDNF表达下降，逍遥散可改善以上异常，而给予BDNF抗体后，逍遥散的作用被抑制。以上结果提示逍遥散可通过促进卵巢局部调控因子BDNF改善慢性应激所致卵泡发育异常。科学意义：研究阐明了逍遥散对慢性应激肝郁证卵泡发育障碍的作用及其卵巢局部BDNF调控机制，为慢性应激相关女性心身疾病的中医药治疗提供现代生物学理论基础和新思路。