电针治疗卵巢过度刺激综合征的最优参数筛选和机制研究

Ovarian hyperstimulation syndrome (OHSS) is a common complication of controlled ovarian stimulation in assisted reproductive cycles. The main pathophysiology feature of OHSS is the increased vascular permeability and resulting shift of fluids into the third space, which brings great harm even the death to patients. Although it has been a hot research topic in the reproductive medicine field worldwide，there are no generally accepted standards for prevention and control. Our initial study shows great promise with electroacupuncture treatment on the OHSS when the Sanyinjiao (SP6) and Guanyuan (CV4) acupoints were stimulated on a rat model. However the physiology and molecular biology mechanism and the optimal stimulation parameter are still unclear. The objective of our further study is to uncover the mechanism and optimal parameter of electroacupuncture therapy on OHSS. To achieve this, an OHSS rat model will be induced. SP6 and CV4 will be stimulated by acupuncture parameters combined with two factors with 3 levels each, stimulating intensity(light, medium and heavy)and courses(short, medium and long). The effects of electroacupuncture with different parameters on peritoneal and ovarian capillary permeability will be investigated by ort hogonal test. Using the selected optimum stimulation parameters, we will observe the expression of inflammatory cytokines in peritoneal fluid and serum, VEGF and its receptors in ovary and uterus, the changes of vascular endothelial cadherin and claudin. Our study will uncover the mechanism of electroacupuncture on the molecular level, in terms of the anti-inflammatory role and regulate VEGF signaling pathway. Our study will also prove for the first time that electroacupuncture is an efficient therapy for OHSS, thus provides a safe, effective and simple treatment method of traditional Chinese medicine for clinical practice.

卵巢过度刺激综合征（OHSS）是辅助生殖技术促排卵过程中的常见并发症，主要表现为血管通透性增加，重者威胁生命，是国内外生殖医学界的研究热点，但迄今无公认的防治技术。我们前期的初步研究发现电针关元、三阴交穴对治疗OHSS模型鼠有效。但何为最佳刺激参数？具体的生理学和分子生物学机制如何？未做进一步研究。为此，本研究通过以轻、中、重的不同刺激强度，长、中、短的不同疗程，针刺OHSS模型鼠关元、三阴交穴，并以腹膜、卵巢血管通透性为效应指标，采用正交设计方法筛选出最优针刺参数。选此最佳参数组合，通过对比观察腹腔冲洗液、血清的特定炎性因子，卵巢、子宫组织VEGF及受体的表达，血管内皮细胞紧密连接蛋白和钙粘蛋白的变化，揭示电针通过抗炎途径和调控VEGF通路治疗OHSS的作用机制。实验目标的实现，将首次明确电针对OHSS的治疗作用，为临床治疗OHSS引入一个安全、有效、简便的中医治疗手段。

为了研究电针（EA）防治卵巢过度刺激综合征（OHSS）的最佳刺激参数以及内在的分子生物学机制，我们首先通过大鼠模型验证了电针对OHSS的确切治疗效果；再以腹膜、卵巢血管通透性为效应指标，通过不同的分组筛选出了最优的刺激参数：预处理长疗程；接着选此最佳参数干预OHSS模型大鼠，发现电针显著缩小模型鼠卵巢体积、减轻卵巢重量，减少卵巢内黄体数目，通过观察血清激素水平、卵巢、血清特定炎性因子，卵巢血管内皮生长因子（VEGF）的表达，初步探索了电针通过影响内分泌激素水平从而抑制炎症因子分泌、调节VEGF表达，最终遏制OHSS进程的可能机制。我们通过进一步的研究发现，EA显著上调卵巢内交感神经因子NPY的表达，通过与前列腺素类物质合成限速酶COX-2相互作用，减少卵巢促黄体生成因子PGE2的生成，增加溶黄体因子PGF2α与PGE2的比值；同时，调控卵巢内细胞增殖相关分子PCNA的分布与表达,上调细胞周期抑制分子p27,发挥其抑制黄体生成并促进黄体结构性与功能性退化的作用,进而影响卵巢类固醇激素合成通路（FSHR(LHR)/PKA/CREB）的活性，下调类固醇激素合成相关酶StAR、Cyp11a1、17β-HSD、Cyp19a1 mRNA及蛋白的表达，从而降低了血清激素水平。我们首次在动物模型中发现了EA影响卵巢功能治疗OHSS的证据，为电针在辅助生殖技术中的临床应用提供了分子生物学依据，这为ART过程中治疗OHSS提供了一个崭新的有效措施。