BDNF在经皮穴位电刺激改善IVF妊娠结局中的作用及机制研究

Treatment infertility with acupuncture has a long history, which was proved to be safe and effective. Many reports had showed that using acupuncture could improve the pregnancy outcomes of in vitro fertilization-embryo implantation (IVF-ET), however, the detailed mechanisms underlying which were still unknown. Both the brain derived neurotrophic factor and its receptors were found in the key brain areas of hypothalamic-pituitary-ovarian axis. It indicated that BDNF might be related with the reproductive ability of the female. Previous reports had showed that the quantity of BDNF in the hypothalamus and other related brain areas increased significantly after the treatment of acupuncture. Moreover, our recent studies found that the BDNF in follicular fluid also increased significantly for those women who received acupuncture during IVF treatment. All those results indicated that BDNF might be involved in the increased pregnant rates of IVF-ET induced by acupuncture. In order to prove this hypothesis, on the basis of our previous studies, we will use BDNF met variant carriers and BDNF genetically modified mice as the research objects, with the help of a variety of modern techniques, to observe the changes of those indicators related with reproduction during the combined treatment of TEAS and IVF-ET, with the quality of BDNF in different state. Through this study, it will help us clarify the mechanism underlying the improved pregnancy outcomes of IVF by TEAS and provide theory support for the application of TEAS in clinical therapy.

针灸治疗不孕症历史悠久，安全有效。在体外受精-胚胎移植（IVF-ET）过程中施加针灸治疗以提高妊娠率的研究已大量开展，虽效果较好，但迄今为止其作用机制仍然不清。脑源性神经营养因子（BDNF）及其受体在下丘脑-垂体-卵巢轴中都有分布，与女性生殖功能相关。研究表明，针灸治疗可以显著提高下丘脑等部位的BDNF含量。我们前期工作也发现，接受IVF治疗的患者配合经皮穴位电刺激（TEAS）干预后卵泡液内BDNF含量明显增多，提示BDNF可能参与调控针灸改善IVF妊娠结局的作用。在此基础上，本项目将以BDNFmet基因变异携带者及两种BDNF转基因小鼠为研究对象，利用多种现代医学手段，从基础到临床，观察TEAS治疗对BDNF水平的影响，分析BDNF与多项生殖功能指标改变的相关性并探索其可能的作用途径。通过本研究，可进一步认识TEAS改善IVF妊娠结局的机制，为临床治疗相关疾病提供理论支持和实验依据。

针灸治疗不孕症历史悠久，安全有效。在体外受精-胚胎移植（IVF-ET）过程中施加针灸治疗以提高妊娠率的研究已大量开展，虽效果较好，但其作用机制仍然不清。脑源性神经营养因子（BDNF）及其受体在下丘脑-垂体-卵巢轴中都有分布，与女性生殖功能相关。研究表明，针灸治疗可以显著提高下丘脑等部位的BDNF含量。经皮穴位电刺激（TEAS）可重复性好，便于数据输出及统计。本项目分别使用TEAS开展临床及动物两个方面研究。临床方面 收集了1028例行IVF-ET治疗患者，将其随机分为TEAS治疗和安慰刺激治疗两大组。观察和研究了TEAS对体内BDNF含量的影响以及抑制BDNF作用对TEAS改善IVF妊娠结局的影响，结果发现：①.TEAS能够提高BDNFval/val基因型患者月经第3日血清BDNF浓度、HCG日血清BDNF浓度及卵泡液中BDNF浓度；TEAS能够降低BDNFval/val基因型患者的月经第3日的卵巢动脉S/D、PI、RI值；TEAS能够降低BDNFval/val基因型患者月经第3日血清FSH水平，提升血清E2水平。②.TEAS能够减少BDNFval/val基因型患者在IVF过程中Gn用量，提高获卵数、受精率、优胚率和胚胎种植率。③.BDNFmet变异基因能够抑制TEAS调控BDNF浓度及改善IVF妊娠结局的作用。含有BDNFmet变异基因的患者血清及卵泡液中BDNF含量明显低于正常人，且在IVF中使用更多的Gn量，获卵数、受精率、优胚率及胚胎种植率更低。④.IVF患者月经第3日卵巢动脉S/D、PI、RI值与月经第3日血清BDNF浓度呈负相关；月经第3日血清FSH水平与血清BDNF浓度呈负相关，第3日血清E2水平与血清BDNF浓度呈正相关；获卵数、受精率、优胚率与卵泡液中BDNF浓度呈正相关。动物研究以BDNF+/-小鼠为研究对象展开。我们分别对BDNF+/-及同窝wild type小鼠进行TEAS或安慰刺激三个动情周期后，分别给与孕马血清促性腺激素模拟人类IVF过程进行超排卵刺激。最终发现：TEAS在野生型小鼠部分生殖环节中的积极作用，包括基础雌二醇的升高、窦卵泡数目增加以及促排卵治疗后卵子成熟率的提高。但同样也观察到BDNF表达的缺失会抑制TEAS改善小鼠生殖的作用。