TLR4通过调控子宫内膜间质细胞生物学行为介导子宫内膜异位症发病的分子机制

It's well known inflammation plays an important roles in the pathogenesis of endometirosis.Compared to normal endometrium ,eutopic endometrium of patients with endometriosis contains more higher capacity of adhesion and invasion. When refluxed into the peritoneal cavity eutopic endometrium debris was more easily to implant at ectopic sites, and Our present studies showed that the expression of IL-8/CXCR1、AKT and β-catenin in eutopic endometrium was strongly than that of normal endometrium. IL-8/CXCR1 are involved in the pathogenensis of endometriosis by up-regulating proliferation and growth and restricting apoptosis in endometrial stromal cells (ESCs) by activating the PTEN/AKT pathway and mediating the expression of survivin and Bcl-2. The upstream regulatory molecule and its mechanism of the secretion of IL-8 peritoneal millieu is abscure. In this study we will build cell co-culture system of ESC, EM42(cell line of endometrial epithelial) and HMrSV5 (cell line of HPMC) to imitate the peritoneal microenvironment of patients with EMT and to evaluate the regulatary mechanism of TLR4 to the secretion of IL-8 and IL-8/CXCR1 to the biological behavior of ESC. We will build animal model to test and verify the role of inflammation in the pathogenesis of EMT. Our study will throw a light in the pathogenesis of the iniation and development of EMT in the inflammatory millieu.

腹腔局部的炎症反应在内异症发病过程中起重要作用，相对于正常内膜在位内膜的粘附和侵袭力更强，但炎症在其中的调控机制不详。我们前期研究发现，与正常内膜相比，在位内膜高表达IL-8/CXCR1、AKT及β-catenin。在位子宫内膜间质细胞（ESC）可通过活化PTEN/AKT信号通路、上调Survivin和Bcl-2表达促进ESC增殖，抑制其凋亡。但腹腔微环境中IL-8的上游调控信号及其对ESC粘附及侵袭的调控机制尚不明确。本课题将构建ESC、腺上皮细胞系及腹膜间皮细胞系的细胞共培养体系，模拟腹腔微环境，并分别从组织、细胞及分子水平探讨上游调控信号TLR4对IL-8调控的分子机制及IL-8/CXCR1对ESC生物学行为调节作用及其分子机制，并通过动物实验进行体外验证，解析炎症对腹腔微环境子宫内膜异位症发病的作用。本课题的完成将为炎症微环境下异位内膜的形成及进展提供新的实验模式和研究思路。

腹腔局部的炎症反应在内异症发病过程中起重要作用，我们前期研究发现，与正常内膜相比，在位内膜高表达IL-8/CXCR1、AKT及β-catenin。在位子宫内膜间质细胞（ESC）可通过活化PTEN/AKT信号通路、上调Survivin和Bcl-2表达促进ESC增殖，抑制其凋亡。为进一步解析腹腔微环境中IL-8的上游调控信号及其对ESC粘附及侵袭的调控机制,本项目构建了ESC、EM42（子宫内膜腺上皮细胞系）及腹膜间皮细胞系共培养体系，研究异位灶微环境中TLR4对IL-8/CXCR1分泌的调控作用，通过实验证实了ESC、EM42及HMrSV5 三种非免疫细胞共培养体系可上调IL-8及其配体CXCR1表达，U937细胞及LPS、TNF-α、PGE2等炎性因子可上调IL-8及其配体CXCR1表达，TLR4中和性抗体可抵抗共培养体系IL-8/CXCR1表达。此外，本项目构还初步建立了SD大鼠EMT动物模型。观察到子宫内膜异位病灶在大网膜、肝脏及胰腺的部位生长，病理切片可见移植物中包含有子宫内膜上皮细胞，腺体和间质的生长，并有分泌活动。为在动物水平上进一步验证TLR4通过促进腹腔微环境中IL-8分泌调控子宫内膜间质细胞生物学行为的分子机制奠定了基础。本项目分别从组织、细胞及分子水平探讨上游调控信号TLR4对IL-8调控的分子机制及IL-8/CXCR1对ESC生物学行为调节作用及其分子机制，解析炎症对腹腔微环境子宫内膜异位症发病的作用。.通过近几年的科学实验，基本验证了科学假说，同时完成了4篇SCI论文的撰写及发表，参与培养硕士3名，顺利完成本科研项目。