基于肿瘤血管芽生调控网络剖析丹参酚酸/丹参酮的整合效应

Danshen is a most frequently used medicine for treating tumors in Traditional Chinese Medicine. However, it is still uncertain whether Danshen promotes tumor metastasis or inhibits it, complicating its clinical use. With the aid of several NSFC programs, our group has demonstrated that Danshensu and Cryptotanshinone, two major components in Danshen, inhibit tumor angiogenesis, but little is known about the underlying mechanisms. The current program puts forward the approach of “Network construction-Drug target identification-Model validation”, and will use the approaches of bioinformatics and network topology coupled with LARS algorithm to establish the regulatory network of tumor angiogenic sprouting. We will use transgenic fluorescence zebrafish tumor vascular sprouting model to validate the critical role of Ephrin-B2 and VEGFR-3 in modulation of vascular sprouting via forward and reverse genetic approaches. We will further use the established 3D dynamic and visible vascular sprouting models and the 3D vascular sprouting models based on co-culture of tumor cells and endothelial cells to study the contributions of Danshen effective component groups that affect key targets to Danshen effects by protein molecular docking technology,directional knockout and quantitative pharmacology methods, showing that Danshen effective component groups exert their integrated effects on tumor vascular sprouting regulatory network via multiple components and multiple targets. These studies will provide novel approaches and strategies for the study of multiple components and multiple targets of traditional Chinese medicines.

丹参在中医治疗肿瘤的方药中，出现的频率极高，但对肿瘤转移是促进还是抑制报道不一，给临床医生带来困惑。课题组在以往多个国家自然基金项目的资助下，发现丹参中丹参素和隐丹参酮对肿瘤血管生成有抑制作用，但分子机制不明。本课题提出“网络构建-药靶识别-模型验证”的思路，借助生物信息学、网络拓扑学结合LARS算法构建肿瘤血管芽生调控网络，利用转基因荧光斑马鱼肿瘤血管生成模型通过正向和反向遗传学手段证实Ephrin-B2和VEGFR-3在调控肿瘤血管芽生中起到关键作用，同时运用实验室已建立的三维、动态、可视化的体外3D血管芽生模型和肿瘤细胞-内皮细胞三维共培养血管芽生模型，采取蛋白分子对接技术、定向剔除技术和定量药理学的策略研究影响关键靶标的丹参活性成分群在丹参中的贡献与地位，明确丹参活性成分群对肿瘤血管芽生调控网络的不同环节通过多靶点、多途径经整合而发挥作用。为中药多成分多靶点的研究提供新思路。

丹参在中医治疗肿瘤的方药中，出现的频率极高，但对肿瘤转移是促进还是抑制报道不一，给临床医生带来困惑。丹参活性成分主要分为两类：脂溶性的丹参酮类和水溶性的丹参酚酸类化合物，脂溶性的丹参酮类成分包括丹参酮Ⅰ、丹参酮ⅡA、丹参酮ⅡB、隐丹参酮等，水溶性的总丹酚酸类成分包括丹参素、咖啡酸、丹酚酸A、丹酚酸B等。项目组总结了丹参中不同成分的作用特点，实验结果发现丹参素、丹酚酸A、丹酚酸B具有抑制体内肿瘤转移的作用，具体表现在抑制VEGF诱导的管腔结构形成及鸡胚尿囊膜血管生成数等；隐丹参酮具有抑制VEGF诱导的大鼠动脉环血管芽生及斑马鱼肿瘤血管芽生的作用。鉴于肿瘤血管生成是一个复杂的网络调控过程，我们推测丹参活性成分群有可能通过作用于关键靶标来调控血管芽生网络而产生抑制肿瘤血管生成的终末效应。项目组提出“网络构建-药靶识别-模型验证”的思路，借助生物信息学、网络拓扑学结合LARS算法构建肿瘤血管芽生调控网络并发现了调控网络中主要靶点TNF-α、VEGF、VEGFR-2、VEGFR-3、Dll4、Notch1、Ephrin-B2，运用转基因荧光斑马鱼肿瘤血管生成模型及血管芽生模型证实了VEGFR-3、Ephrin-B2、TNF-α等关键靶点在调控肿瘤血管芽生中起到的关键作用，采取蛋白分子对接技术、定向剔除技术和定量药理学的策略研究影响关键靶标的丹参活性成分群在丹参中的贡献与地位，明确丹参活性成分对肿瘤血管芽生调控网络的不同环节通过多靶点、多途径经整合而发挥作用。为中药多成分多靶点的研究提供新思路。