基于水通道蛋白的少腹逐瘀汤有效组分干预子宫内膜异位症“粘附-侵袭-血管生成”机制研究

Endometriosis (EMs) has some characteristics of similar and widespread planting, growing, invasive with malignant tumors, and clinical manifestations of dysmenorrhea, infertility, high recurrence rate, seriously affects the patient's life quality of childbearing age and reproductive health. Shaofuzhuyu Tang (SF) is a classic recipe for the treatment of cold blood stasis syndrome.On the basis of pre-clinical and experimental studies, it had shown that: SF could produce a therapeutic effect on EMs by interventing the expression levels of relevant indicators of EMs.This project, based on previous research, guiding the traditional Chinese medicine diagnosis and treatment with disease of governance theory, as aquaporin (AQP) as a starting point, use of disease and syndrome model, determinate the AQPs expression of tissue, peritoneal fluid and cells of EMs by RT-PCR and Western Blot methods; the relevance research of adhesion - aggression- angiogenesis through cell adhesion, scratch, transwell test and VEGF expression and microvessel density of EMs, and use of siRNA experiment to verify; at the same time through the relationship between hemorheological indexes, to explore the relevance between AQPs and cold blood stasis syndrome, and thus systematically to explain the formation mechanism of SF intervention EMs, and provide a scientific basis for medical treatment for EMs.

子宫内膜异位症（EMs）具有与恶性肿瘤相似的广泛种植、生长、浸润特性，临床表现为痛经、不孕等，且复发率高，严重影响育龄患者的生活质量和健康。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可以通过干预EMs形成相关指标的表达而达到对EMs的治疗作用。本项目在前期研究基础上，以中医药辨证论治与辨病论治理论为指导，以水通道蛋白（AQP）为切入点，采用病证结合模型，通过RT-PCR和Western Blot方法测定EMs组织、腹腔液及细胞中AQPs表达；通过细胞黏附、划痕、transwell试验及VEGF表达和EMs微血管密度变化，研究与EMs“粘附-侵袭-血管生成”的相关性，并采用siRNA干扰试验进行验证；同时通过分析与血液流变学各项指标的关系，探讨AQPs与中医血瘀证的相关性，进而系统阐释SF干预EMs形成的作用机制，为中医药治疗EMs提供科学依据。

子宫内膜异位症（EMs）具有与恶性肿瘤相似的广泛种植、生长、浸润特性，临床表现为痛经、不孕等，且复发率高，严重影响育龄患者的生活质量和健康。少腹逐瘀汤为治疗寒凝血瘀型EMs的经典方剂，本项目在前期研究基础上，首先，以中药血清药物化学理论为指导，采用UPLC/MS方法对少腹逐瘀汤进行体外全成分分析基础上，明确了少腹逐瘀汤治疗寒凝血瘀型EMs的体内主要有效成分，并进一步以指纹图谱结合主要活性成分指标建立了质量控制方法。同时，以中医药辨证论治与辨病论治理论为指导，以自体移植结合冰水浴法建立了寒凝血瘀型EMs病证结合模型，采用HE染色、RT-PCR、Western Blot等方法对病证结合模型大鼠研究结果，结果显示，寒凝血瘀型EMs大鼠伴随水通道蛋白AQP1、AQP2、AQP5、AQP8各亚型以及AKT1表达异常，揭示了水通道蛋白与寒凝血瘀型EMs的内在联系；以少腹逐瘀汤及其主要代表性活性成分分别对寒凝血瘀型EMs模型大鼠进行干预，在通过药理等指标证实药物对寒凝血瘀型EMs治疗有效的基础上，HE染色、RT-PCR、Western Blot等研究结果均表明，少腹逐瘀汤及主要活性成分对水通道蛋白AQP1、AQP2、AQP5、AQP8各亚型均有不同程度的回调作用，阐明了对水通道蛋白的调节为少腹逐瘀汤治疗寒凝血瘀型EMs的作用机制之一。研究为EMs的发病机制阐释及中医药治疗EMs提供新思路。