甘遂半夏汤治疗肝癌和缓解肝癌腹水并发症的药效物质及作用机制研究

甘遂半夏汤为张仲景《金匮要略》经方之一，治疗肝癌及缓解肝癌腹水并发症疗效确切，具“攻癌毒”、“克腹水”的多层次、多靶点、标本兼顾的明显优势，但功效成分及作用机理尚不明确。申请者在前期工作中，发现甘遂半夏汤提取物具有明显的体内抗肝癌活性(醇提物>水提物)，还能显著降低荷瘤小鼠腹水量和增加尿量(水提物>醇提物)。初步的作用机理研究表明，组方对肿瘤及脾脏免疫细胞具调节作用，能降低肿瘤相关的炎性损伤，还可抑制PI3K-Akt-mTOR和MAPK/ERK信号通路相关蛋白的表达。申请者拟以组方水提物和醇提物为研究对象，从组方整体出发，采取从整体到部分、逐层递进的策略，以活性筛选和UPLC-QTOF-MS为指导，寻找“标本兼顾”的高活性成分或组分，确定发挥作用的原型成分和入血成分；通过中药化学-血清化学-药理学的联动分析，阐明组方的核心药效物质基础和作用机制，为中药复方的研究提供新的思路和模式。

Gansui-Banxia decoction (GSBXD), as a classic TCM formula from Chinese medical sage Zhang Zhongjing, has been used for the treatment of hepatocellular carcinoma (HCC) and malignant ascites. It has obvious advantages in treating HCC and malignant ascites, not only "anti-cancer" but also “controls ascites" with multi-component and multi-target agents that essentially achieve therapeutic effectiveness. However, its effective ingredients, blood-entry ingredients, and the pharmacological mechanism of diarrhea, water-expelling and anti-tumor have not been fully elucidated. . In the previous study, we have found that the extract of GSBXD can potently exhibit anti-cancer activities in vivo (ethanol extract > water extract), and significantly inhibited the ascites volume and increase urine volume of the tumor-bearing mice (water extract > ethanol extract). Further mechanism studies showed that they could regulate the expression of tumor and spleen immune cells, and inhibit the expression of PI3K-Akt-mTOR and MAPK/ERK signaling pathway related proteins, speculating that the anti-tumor effect is related to the signaling pathways mediated by EGFR, FGFR and VEGFR receptors. . Therefore, taking the water extract and alcohol extract of GSBXD as the research object, the project is designed to isolate and identify the effective constituents, the active prototype effective components and blood-entry ingredients of GSBXD, by using various of modern chromatography and NMR technologies, under the guidance of UPLC-QTOF-MS and bioactive assay. The antitumor and inhibition of cancerous ascites potential of the active compounds or extracts will be comprehensively evaluated from the aspects of cell proliferation, cell migration, cell cycle and cell apoptosis, and regulation of spleen and tumor immune cells. Meanwhile, the prohibitive effect of the potent compounds on the anti-apoptotic PI3K/AKT signaling pathway was further investigated, combining the pharmacological studies of anti-HCC and malignant ascites in vivo and in vitro. The possible anti-HCC and cancerous ascites mechanism of the active compounds will be explored to identify the potential target. . This research may develop a comprehensive system for understanding the pharmacological mechanisms of GSBXD acting on HCC and malignant ascites, and further clarify and expand the application scope of GSBXD, to promote the development and application of new drugs in the field of HCC and malignant ascites. It also will provide new ideas for the study of traditional Chinese medicine.