补肾法、疏肝法改善子宫内膜容受性与调控VEGFR-2血管生成相关信号通路的关系

The incidence of infertility is up to 7%-10% in our country, and poor endometrial receptivity mainly accounts for implantation failure.Thus improving endometrial receptivity(ER) become a hot topic at home and abroad. Previous studies showed that kidney-invigorating and liver-soothing treatments could increase endometrial thickness, regulate reproductive hormones and their receptors, enhance cell adhesion, control endometrial VEGF and its receptor, which suggesting that the two treatments have the effect of improving ER. VEGF is a major regulator of angiogenesis and its receptor VEGFR-2 mediated PI3K, MAPK pathway is involved in angiogenesis. In the research, we will explore the role of two treatments in improving ER from endometrial morphology. Meanwhile, we planed to investigate the effects of two treatments on VEGFR-2, its downstream molecular consequences of PI3K, MAPK pathway, PCNA, cyclin D1, and MMP9 in endometrium from implantation failure patients, in human vascular endothelial cells and in super-ovulation mice bed endometrium. We will examine the relationship between two treatments in improving ER and regulating VEGFR-2 related signaling pathway in angiogenesis. This study plays an important role in eluciating the mechanism of menstruation and bearing, enriching the traditional Chinese medicine theory of "Kidney dominating reproduction", "for women liver is congenital", and improving reproductive theory of Traditional Chinese Medicine.

我国不孕症发病率为7%-10%。着床失败原因主要在于内膜容受不良。因此，改善子宫内膜容受性(ER)成为国内外研究热点。前期研究发现补肾法、疏肝法可增加子宫内膜厚度、调节生殖激素及其受体、提高细胞黏附力、调控内膜VEGF及其受体，提示两法具有改善ER的作用。子宫内膜血管生成是胚胎着床的关键。VEGF是最主要的血管生成调节因子，其受体VEGFR-2所介导的PI3K、MAPK通路参与血管生成。本项目拟从形态学方面研究两法改善ER的作用，并通过两法对着床失败患者子宫内膜、血管内皮细胞、超促排卵小鼠围着床期子宫内膜VEGFR-2及其血管生成通路以及促血管生成基因PCNA、cyclin D1、MMP9的影响，研究两法改善ER与VEGFR-2血管生成相关信号通路的关系，比较两法作用环节之异同。这对于阐释中医调经种子作用机制、丰富"肾主生殖"、"女子以肝为先天"理论内涵、完善中医生殖理论具有重要的意义。

我国不孕症发病率为 7%-10%。体外受精-胚胎移植（IVF-ET）是目前治疗不孕症的主要手段之一，但妊娠率仍徘徊在30%-40%，着床失败原因主要在于内膜容受不良。因此，改善子宫内膜容受性(ER)成为国内外研究热点。近年研究发现，促进子宫内膜血管新生是改善子宫内膜容受性的关键，是提高IVF-ET妊娠率的有效途径。VEGF是最主要的血管生成调节因子，其受体VEGFR-2所介导的PI3K、MAPK通路参与血管生成。本课题组在前期证实补肾法、疏肝法可改善子宫内膜容受性的基础上，根据中医“肾主生殖”、 “女子以肝为先天”的理论，进行补肾法、疏肝法改善子宫内膜容受性的比较研究，从临床、动物、细胞水平证实补肾法、疏肝法通过提高血清雌孕激素水平，上调子宫内膜ER、PR的表达，改善子宫内膜形态、胞饮突形态和数量，从而提高子宫内膜容受性，但两法之间差异不明显。同时进一步研究补肾法和疏肝法改善子宫内膜容受性的分子机制，发现两法可影响着床失败患者子宫内膜、血管内皮细胞、超促排卵小鼠围着床期子宫内膜的VEGFR-2、血管生成通路（MAPK和Akt信号通路）以及促血管生成基因PCNA、cyclin D1、MMP9和通透相关指标eNOS，促进内皮细胞的增殖、迁移和通透性，进而促进子宫内膜血管生成。然而，两法作用环节有所不同，补肾法通过激活ERK信号通路促进PCNA表达，激活ERK、JNK、P38信号通路促进Cyclin D1表达，激活JNK、P38信号通路促进MMP9的表达；疏肝法通过P38信号通路促进PCNA和MMP9的表达，通过JNK、P38促进Cyclin D1表达。补肾法、疏肝法也可激活PI3K/Akt信号通路和eNOS，增加血管通透性，进而改善子宫内膜容受性，提高妊娠率。补肾法在促进血管增殖方面优于疏肝法，在促进血管迁移、增加血管通透性方面，疏肝法优于补肾法。这对于阐释中医调经种子作用机制、完善中医生殖理论具有重要的意义。