“三棱-莪术”组分配伍干预子宫肌瘤细胞增殖的基质成纤维细胞活化机制研究

Uterine Leiomyoma(UL)is the most common benign tumors of women,Which is belonged to “Zheng jia” “ji ju” in TCM,TCM has the advantage and the disease incidence is rising,So Sanleng-Ezhu as the compatibility is commonly used in clinical and enhance efficacy each other.The former NSFC(N0.81001668) study shows that uterine leiomyoma cells is hypertrophy, disarray and focal hyperplasia,and San-leng e zhu compatibility significantly inhibit proliferation of leiomyoma cells, but the mechanism is unclear. Studies have shown that stromal fibroblasts (Fib) activation plays an important role in hair growth in ULC,so we put forward the “sanlneg-ezhu compatibility to inhibit leiomyoma cells proliferation by interfering with fib Activation” hypothesis.This topic is to deepen the preliminary study,We will Closely combine the basic and clinical in this project, adopt two model in vivo and in vitro(Human cell model of UL and UL ras ),from the perspective of component compatibility and Fib activation,We will adopt modern science and technology researching Fib activation markers,ECM, growth factors, proliferation activity and associated signaling molecules, multi-level, multi-angle to explore the mechanism of San leng-Ezhu compatibility   inhibiting   leiomyoma cell proliferation, which is important to elucidate Sanleng-ezhu compatibility law, prevent and treat of ULmechanism, find new targets, and provide the basis for the clinical application and new ideas.At the same time it will provide a demonstration of drug compatibility ,further validate the academic value of the classical TCM.

子宫肌瘤是女性最常见良性肿瘤，发病呈上升趋势，属中医学“癥瘕”、“积聚”范畴,中医药治疗有优势，临床常用三棱莪术相须配伍。前期国家自然基金研究发现肌瘤细胞肥大、排列紊乱、局灶性增生，三棱莪术组分配伍能明显抑制肌瘤细胞的增殖，但作用机制不清楚。已有研究表明基质成纤维细胞(Fib) 活化在肌瘤生发中起重要作用，我们提出“三棱莪术组分配伍通过干预Fib活化来抑制肌瘤细胞增殖”假说。本课题是前期研究的深化，结合基础与临床，采用体内外模型（人子宫肌瘤细胞模型和大鼠子宫肌瘤模型），从组分配伍及Fib活化角度，应用现代科学技术研究Fib活化标记物、ECM、生长因子、增殖活力及相关信号分子，多层次、多角度探索三棱莪术组分配伍干预肌瘤细胞增殖作用机制，对阐明三棱莪术配伍规律及防治子宫肌瘤机制、寻找新靶点具有重要意义，为临床应用提供机制依据和新思路，为药对配伍研究提供示范，进一步验证祖国医学经典理论的学术价值

子宫肌瘤是女性最常见良性肿瘤，发病呈上升趋势，属中医学“癥瘕”、“积聚”范畴, 中医临床常用三棱莪术相须配伍。前期国家自然基金研究发现三棱莪术组分配伍能明显抑制肌瘤细胞的增殖，但作用机制不清楚。本项目在前期研究基础之上，从体内体外两方面研究“三棱-莪术”组分配伍干预子宫肌瘤细胞增殖的基质成纤维细胞的活化机制。采取雌孕激素负荷法能够成功构建子宫肌瘤动物模型，研究三棱-莪术组分配伍抑制大鼠子宫肌瘤成纤维细胞活化机制，观察子宫组织病理学改变，检测大鼠血清E2、P水平和大鼠子宫组织ER、PR、FAP、CollagenⅠ、TGF-β、Fibronectin（FN）、AKT、ERK1/2、MEK表达情况。经人原代细胞培养和纯化后，得到了高纯度的成纤维细胞，检测各组成纤维细胞增殖活力、FAP、ER、PR、TGF-β、CollagenⅠ、Laminin、p-AKT、p-MEK、p-ERK1/2、FN、AKT、MEK、ERK1/2表达差异。. 结果发现使用雌孕激素负荷结合外因素刺激法可成功建立气滞血瘀型子宫肌瘤大鼠模型；三棱总黄酮-莪术挥发油配伍可降低气滞血瘀型子宫肌瘤大鼠子宫中TGF-β1含量，抑制子宫肌瘤成纤维活化和TAF活化以及p-AKT、p-ERK1/2、p-MEK及Collagen I表达。证实在肌瘤相关成纤维细胞处于活化状态，并高表达 FAP、ER、PR、 TGF-β，并能分泌 ECM 相关蛋白。“三棱-莪术”组分配伍可抑制肌瘤相关成纤维细胞活化，降低 FAP、 ER、PR、TGF-β、p-AKT、p-MEK、p-ERK1/2 及 ECM 的表达，且“三棱-莪术”组分配伍的作用总体要优于三棱总黄酮与莪术挥发油单组分。说明“三棱-莪术”组分配伍可通过干预子宫肌瘤细胞增殖的基质成纤维细胞活化机制达到防治子宫肌瘤的目的。. 本课题是前期研究的深化，结合基础与临床，采用体内外模型（人子宫肌瘤细胞模型和大鼠子宫肌瘤模型），从组分配伍及Fib活化角度，应用现代科学技术研究Fib活化标记物、ECM、生长因子、增殖活力及相关信号分子，多层次、多角度探索三棱莪术组分配伍干预肌瘤细胞增殖作用机制，对阐明三棱莪术配伍规律及防治子宫肌瘤机制、寻找新靶点具有重要意义，为临床应用提供机制依据和新思路，为药对配伍研究提供示范。