消癌解毒方调控炎癌转化的活性组分及配伍机制研究

Xiaoai jiedu recipe is an effective recipe prescribed for cancer treatment formed by compatibly combining herbals through treatment based on pathogenesis differentiation, directed by the “ai du” pathogenesis theory. Previous studies showed that xiaoai jiedu recipe played a key role in cancer treatment through intervening tumor and inflammatory microenvironments. The mechanism of inflammation-cancer transformation is closely associated with tumor microenvironment, especially with inflammatory microenvironment. Our study suggests that xiaoai jiedu recipe can prevent and treat cancer by regulating and controlling inflammation-cancer transformation network. This project aims to investigate the effects of xiaoai jiedu recipe, its related components and parts, and invidiual components on colon inflammation-cancer transformation model and key signaling pathways of NF-κB, STAT3, as well as to analyze and isolate active components, through in vivo and in vitro experiments, combination of chemical separation and analysis and pharmacological efficacy indicators, application of modern chromatographic spectrum technology, by means of systems biology, molecular biology, cell biology, epigenetics and the like, on macroscopic and microscopic levels of overall, cells, genes and molecules. With the aid of modern spectroscopy in series technology, we explored the active components regulating and controlling inflammation-cancer transformation in the recipe and molecular mechanism thereof, and reveal compatibility mechanism of the recipe, laying a foundation for prevention and treatment of cancer and formulation of anti-cancer drugs.

消癌解毒方是基于中医肿瘤癌毒病机理论，通过辨治癌毒病机，进而配伍形成的抗肿瘤临床有效验方。前期研究表明消癌解毒方干预肿瘤微环境及炎性微环境发挥抗肿瘤作用，而炎癌转化机制与肿瘤微环境，特别是炎性微环境关系密切。我们研究认为，消癌解毒方可能通过调控炎癌转化网络防治肿瘤。本项目结合文献研究及前期实验基础，以癌毒病机理论为指导，通过体内及体外实验，化学分离分析与药理药效指标相结合，应用现代色谱波谱技术，结合系统生物学、分子生物学、细胞生物学、表观遗传学等手段，从宏观与微观，基于整体、细胞、分子基因等多层次，观察分析消癌解毒方、相关组分及部位、单体成分对结肠炎癌转化模型及其关键信号通路NF-κB、STAT3的影响，分析分离活性组分，重点辅以现代谱学串联技术，探讨复方调控炎癌转化的活性组分及分子机制，揭示复方配伍机理，为应用癌毒病机理论指导肿瘤防治以及抗肿瘤药物的创制提供依据。

本研究以消癌解毒方为研究对象，以癌毒病机理论为指导，遵循转化医学研究思路，首先对消癌解毒方的有效成分及体内代谢过程进行研究。鉴定了102个组分，其中黄酮类化合物30个，三萜类化合物19个，有机酸12个，甾体皂苷9个，环肽9个，菲类化合物7个，氨基酸5个，生物碱3个，其它化合物8个。通过分析口服复方后大鼠血浆和尿液中的代谢物，共鉴定出70种化合物，包括15种主要成分和55种代谢物，并对氢化、羟基化、甲基化、磺化和葡萄糖醛酸化等途径进行了评价。通过16S rRNA的测序分析了黄连-乌梅药对与消癌解毒方中生物碱类成分的含量差异，并比较药对与全方对小鼠肠道菌群的影响。基于 TMT 标记的蛋白质组学技术检测消癌解毒方及其组分药干预肝癌大鼠后的肝组织差异表达蛋白，通过建立结肠癌细胞荷瘤小鼠模型及结肠炎相关性结肠癌模型，结合相关体外实验，观察了该方对结直肠癌的抑制作用及对结直肠非可控性炎症发展为癌症病程的干预作用。同时观察消癌解毒方及其各药物组分对结直肠癌模型相关细胞、细胞因子和信号通路等炎癌转化调控网络的影响，重点围绕肿瘤巨噬细胞极化状态的改变及其调控因素对消癌解毒方的作用机制进行了探讨。体外研究结果表明，消癌解毒含药血清通过抑制结直肠癌细胞的糖酵解作用，诱导肿瘤细胞发生周期阻滞，进而诱导细胞凋亡。体内研究结果显示，消癌解毒方及其组分药和有效成分抑制炎-癌转化过程的作用可能是通过抑制肿瘤炎性微环境中的炎性因子的表达，干预了PI3K/Akt、TLR4/NF-κB、JAK2/STAT3及TGF-β通路。实验结果为寻找靶向巨噬细胞的免疫干预的抗肿瘤药物的研究提供了新的思路及理论基础，为应用癌毒病机理论指导中医药防治肿瘤提供科学依据。