基于性激素-关键基因-超微结构网络的少腹逐瘀汤有效组分干预寒凝血瘀证EMs子宫内膜容受性机制研究

Endometriosis ( EMs ) is a major reason for infertility, and the health problem caused by the decrease of pregnancy potential has rised to social problems which seriously influences the quality of life in patients of reproductive age. Shaofu Zhuyu Decoction ( SF ) is a classic prescription for treating blood stasis led by cold accumulation of EMs. Previous clinical and experimental studies show that SF has a definite therapeutic effect on EMs infertility.It has also been indicated that the SF contained serum shows therapetic effect to EMs by affecting the expression level of related indicators of eutopic endometrial cells during the course of Attachment-Aggression-Angiogenesis.Based on the preliminary study, the present research aims for clafication of the pharmacoligical components of SF using EMs cell. The correlation between the effect of EMs on endometrial receptivity and pregnancy rate will be discaused according to the changes of progesterone, estrogen serum level and related receptor expression( hormone),and the changes of endometrial ανβ3 gene and HOXA10 gene ( key gene ), pinopodes ( ultrastructure ), blood rheology changes ( blood stasis ) under the instruction of holistic theory of traditional Chinese medicine taking "sex hormone - key genes - ultrastructure "network as the breakthrough point. The results obtained will help to clarify the mechanism of action of SF on endometriosis infertility, and to provide scientific basis for improvement of the pregnancy rate of EMs by traditional Chinese medicine.

子宫内膜异位（EMs）是不孕症的主要原因，EMs导致的妊娠能力下降，已由健康问题上升至社会问题，严重影响育龄期患者生活质量.少腹逐瘀汤（SF）为治疗寒凝血瘀证EMs的经典方剂，前期临床及实验研究表明：SF治疗EMs不孕疗效确切；SF含药血清通过干预在位内膜细胞"粘附-侵袭-血管生成"相关指标的表达水平而达到对EMs的治疗作用。本项目在前期研究基础上，以SF不同组分为对象，以EMs细胞为工具，筛选其有效组分，以中医药整体观理论为指导，以"性激素-关键基因-超微结构"网络为切入点，采用病证结合模型，从孕激素、雌激素血清含量及相关受体表达变化（性激素）、子宫内膜ανβ3及HOXA10基因（关键基因）、胞饮小泡（超微结构）、血液流变学（血瘀）等方面的变化探讨EMs对子宫内膜容受性的影响及其与妊娠率的相关性，阐释SF治疗子宫内膜异位所致不孕症的作用机制，为中医药提高EMs妊娠率提供科学依据。

子宫内膜异位（EMs）是不孕症的主要原因，严重影响育龄期患者生活质量，少腹逐瘀汤（SF）为治疗寒凝血瘀证EMs的经典方剂，治疗EMs不孕疗效确切，但其作用机制尚不明确。本研究首先以EMs细胞为工具，对少腹逐瘀汤的各个部位进行活性筛选，确定SF有效部位及工艺，采用HPLC方法建立了SF指纹图谱，并同时对其中8个主要成分进行了含量测定，有效控制了方剂质量。采用自体移植结合冰水浴法制备并评价寒凝血瘀证EMs大鼠模型，模型大鼠的受孕率和胚泡着床数均显著低于手术对照组。在此基础上，以中医药整体观理论为指导，以“性激素-关键基因-超微结构”网络为切入点，采用病证结合模型，探讨EMs对子宫内膜容受性的影响及少腹逐瘀汤干预机制：①血液流变学结果表明，EMs血浆及全血粘度均有升高；②激素P、E2及其受体ER、PR研究结果表明，模型组P水平下降、E2上升，受体ER有显著升高，PR变化无统计学意义；③关键基因研究结果显示，基因HOXA-10、干扰素αv、β3的表达均有显著提高；④超微结构：内膜组织中腺细胞、间质基质细胞和上皮细胞的超微结构上均有显著变化；即EMs从以上四方面对子宫内膜容受性产生影响，导致受孕率和胚泡着床数有显著性降低。SF各剂量组对上述四个方面的各指标均有不同程度回调作用，从而改善子宫内膜容受性，达到对EMs不孕的治疗。本研究为中医药提高EMs妊娠率提供了一定的科学依据。