活血化瘀中药独立于抗凝作用外的抗炎-抗肝癌途径分子机制及"扶正"配伍研究

Chinese medicine(CM) prescriptions have demonstrated usefulness in treating cancer. In CM, blood stasis is the key pathogenesis of cancer and promoting blood circulation by removing and blood stasis is the important treatment guideline. Much of studies deal with direct cytotoxic effects or anticoagulation activity, the anticarcinogentic mechanism, unfortunately, is still unclear. On the other hand, studies results indicated that organs deficiency, named "zhengxu" in CM terms, is the basic pathogenesis of cancer. Dangshen (Condonopsis pilosula Radix.) and Jianghuang (Carcumea longa Radix.) are both Chinese herbs for blood activating and stasis-dissolving. Condonopsis saponins and curcumin are the principal bioactive chemicals in Dangshen and Jianghuang, respectively. With the deepening research on pathogenesis of cancer, inflammation and immune microenvironment in situ are considered as the key players in cancer growth, spread and metastasis. Studies have revealed that Chinese herbs for blood activating and stasis-dissolving have powerful anti-inflammatory property. Our previous studies demonstrated that compatibility of Dangshen and Jianghuang plays a important role in supporting slpeen healthy energy, named "fuzheng combination" in TCM terms. These lead to the hypothesis that the anti-hepatoma effecs of blood activating and stasis-dissolving Chinese herbs is based on anti-inflammatory activities independently anti-coagulation effects, in addition, Dangshen and Jianghuang "fuzheng combination" has the synergistic effect to the anti-inflammation and anti-cancer pattern. To prove our hypothesis, we will utilize integrated pharmacodynamic, cellular and molecular methodologies to investigate anti-inflammatory effects, anti-coagulation effects and "fuzheng" synergistic effects of Dangshen, Jianghuang and the combination with the multi-component contrasted mono-component model. The experimental verification of its molecular mechanism will carry out using Treg suppressive function of hepatoma microenvironment, inflammatory TLR4 signaling and effects on angiogenesis. The study results will provide empirical evidence to support the anti-hepatoma mechanism of Dangshen and Jianghuang combinaiton, and lay a substantial foundation for enriching the modern science ideas of "fuzheng".

"血瘀"为癌症的重要中医病机，活血化瘀方药抗癌机制仍未完全明晰，目前多从直接杀伤效应或抗凝角度做出阐释。随着癌症研究的不断深入，炎症微环境状态在肿瘤细胞生长、播散、转移中的重要性越来越受到关注。活血化瘀抗癌中药具有良好的抗炎活性，结合前期基础，我们提出活血化瘀中药独立于抗凝作用外的抗炎-抗肝癌途径假说，并通过系列体外与在体实验从抗凝、抗炎两条途径分析加以验证；进而以免疫、炎症中心环节MDSCs为切入点，观察活血化瘀中药对肝癌微环境的免疫抑制调节、TRL4的炎症促发及血管新生的影响，探讨其分子机制。同时，在中医整体观前提下，根据肝癌的"脾虚"病机，以党参皂苷+姜黄素进行成分配伍，探讨"扶正"配伍对活血化瘀中药抗炎-抗肝癌途径的协同作用机制。本项目的研究，将重新定位，并为建立活血化瘀中药抗癌研究思路，丰富"扶正"的现代科学内涵提供理论支持。

持续的炎症是癌症恶性成长的源泉。根据“血瘀”是肝癌的重要中医病机，以及活血化瘀中药的抗凝与抗炎的双重活性，仅仅从抗凝角度仍很难深入理解和解释活血化瘀中药对肝癌细胞的抑制杀伤作用，以及动物实验中呈现出的抑实体瘤效果。因此我们提出活血化瘀中药独立于抗凝作用外的抗炎-抗肝癌途径假说。为了验证假说，我们进行了如下研究：（1）活血化瘀中药独立于抗凝作用外的抗炎-抗肝癌途径假说验证；（2）以免疫、炎症中心环节MDSCs为切入点，观察活血化瘀中药对肝癌微环境的免疫抑制调节、TRL4的炎症促发及血管新生的影响，探讨其分子机制；（3）以人参皂苷+姜黄素进行成分配伍，探讨“扶正”配伍对活血化瘀中药抗炎-抗肝癌途径的协同作用机制。主要取得了如下成果：（1）理论上验证了活血化瘀中药姜黄素抗炎-抗肝癌的效应途径，细胞及动物实验结果表明，抗凝途径并不是理想的抗肿瘤抗肝癌作用方式；（2）阐明了姜黄素抗炎-抗肝癌途径的分子机制：主要包括抑制诱导MDSCs增殖、募集的因子IL-6、COX-2、PGE2，以及诱导MDSCs活化的通路信号蛋白MyD88、磷酸化IKKs因子的表达，以及降低炎症因子TNF-α、IL-1β的蛋白表达，同时抑制血管新生等方式，改善肝癌的微环境，从而抑制肝癌组织的生长；（3）高剂量的大量使用姜黄提取物，可能会降低姜黄素的抗肿瘤效果，提示活血化瘀中药姜黄所含的多成分，具有一定的内部拮抗效应。（4）明确了扶正中药人参皂苷与姜黄素之间存在协同作用，通过抑制PD-1/PD-L1的蛋白表达，是其重要的抗肿瘤抗肝癌重要机制之一；同时扶正+抗炎的中药抗肿瘤配伍模式，对今后抗肿瘤的中药新药的研发具有指导意义。