补肾活血中药重建子宫内膜异位症免疫微环境抑制盆腔纤维化机制研究

Endometriosis is an estrogen-dependent inflammatory disorder characterized by the presence of endometrial tissue outside the uterine cavity. Pelvic adhesion is an important characteristic of endometriosis, which lead to infertility and pelvic pain. The inflammatory microenvironment of endometriosis give rise to chronic damage of peritoneal tissues and form pelvic fibrosis. Blood stasis is intrinsic pathology of endometriosis. Removing blood stasis is main treating principles for endometriosis. Previous studies had shown that the medicine of nourishing kidney and activating blood treating endometriosis was effective, and it suppressed abdominal adhesion formation. We speculate that pelvic microenvironment of endometriosis may be an outcome of combined action of platelet and macrophages. The activated platelet from repeated bleeding of ectopic endometrial tissue induced the conversion of macrophages to M 2 macrophage phenotype, and lead to abnormal immune inflammatory cascade, the peritoneum tissue damage and pelvic fibrosis. The medicines of nourishing kidney and activating blood prevented and treated pelvic fibrosis of endometriosis by effective intervention on immune inflammatory process. This project intends to explore possible mechanisms about immune microenvironment of endometriosis resulting in chronic injury of peritoneum by establishing cellar co-culture system and adopting the technology of RNA interference etc, and to elucidate the possible mechanism which immune microenvironment of endometriosis induce the pelvic fibrosis by confocal immunofluorescence staining, phosphorylation protein kinase chips with peritoneal mesenchymal cell as the research object. This project are still to clarify the mechanism about preventing and treating pelvic fibrosis of endometriosis with the medicine of nourishing kidney and activating blood by establishing mouse model of endometriosis using syngeneic menstrual endometrial tissue.

子宫内膜异位症（Endometriosis，EM）是雌激素依赖的免疫性炎症疾病。盆腔粘连是EM的重要特征，是导致不孕和盆腔痛的重要原因。EM的炎症微环境造成腹膜组织慢性损伤，形成盆腔纤维化。血瘀是EM的病理状态，活血化瘀为EM基本治疗原则。前期研究显示补肾活血方可有效治疗EM，并抑制腹腔粘连形成。我们推测，EM盆腔微环境形成可能是血小板与巨噬细胞共同作用的结果。EM异位内膜组织反复出血中的活化血小板诱导巨噬细胞呈现M2型改变，导致炎症性免疫反应，引起腹膜组织的持续损伤，促使盆腔纤维化形成；补肾活血中药通过干预EM的免疫炎症过程，防治盆腔纤维化。本项目拟建立细胞共培养体系，探讨产生EM腹膜慢性损伤的免疫微环境的可能机制；以人腹膜间皮细胞为研究对象，探讨EM免疫微环境引起盆腔纤维化的可能机理；用模拟经期内膜组织建立EM小鼠动物模型，从免疫微环境重建角度阐明补肾活血中药防治EM盆腔纤维化的机制。

子宫内膜异位症（Endometriosis，EM）是雌激素依赖的免疫性炎症疾病。盆腔粘连是EM的重要特征，是导致不孕和盆腔痛的重要原因。EM的炎症微环境造成腹膜组织慢性损伤，形成盆腔纤维化。血瘀是EM的病理状态，活血化瘀为EM基本治疗原则。前期研究显示补肾活血方可有效治疗EM，并抑制腹腔粘连形成。我们在本研究中成功构建EM小鼠模型，明确了EM腹腔微环境在疾病进展过程中呈现动态变化特征，早期以炎症反应为主，中后期以损伤修复为主，腹腔粘连和纤维化程度随疾病进展逐渐加重；阐明了EM进展过程中巨噬细胞M1型和M2型极化状态的变化趋势以及相关炎性因子变化特征；完成了补肾活血方对EM模型小鼠盆腔纤维化干预，发现补肾活血方可调控EM模型小鼠腹腔巨噬细胞极化和炎性因子的表达，抑制盆腔粘连和纤维化的发展。在明确补肾活血方调控巨噬细胞极化，改善盆腔微环境，抑制盆腔纤维化的基础上，我们对补肾活血方主要药效成分进行筛选，开展补肾活血方的有效单体姜黄素治疗EM的系列研究，结果表明：（1）通过姜黄素对巨噬细胞极化的调控以及巨噬细胞炎性因子分泌调控的研究，发现姜黄素可以多途径、多靶点调控巨噬细胞极化；（2）通过姜黄素调控巨噬细胞对子宫内膜间质细胞侵袭和凋亡影响的研究，发现EM异位内膜组织存在焦亡，释放大量炎性因子，改变腹腔免疫环境。（3）通过网络药理学分析提示，姜黄素可以良好对接焦亡关键蛋白，并通过抑制氧化应激抑制焦亡治疗EM。我们的研究阐明了免疫微环境在EM中的发病机制；补肾活血方对于临床治疗EM疗效确切；姜黄素是治疗EM的重要候选化合物。