基于炎症微环境NF-κB和MAPK信号通路的新疆软紫草抗肝癌药效物质基础及作用机制研究

Digestive system tumors are called as "inflammation-related tumors", so it exhibits great significance to study the occurrence and development of inflammation-liver cancer from the perspective of "inflammatory and cancer transformation" and its potential mechanism. Previous studies have found that ethanol and hot water extract of Arnebia euchroma showed high anti-HBV activity, and anti-HBV and HIV-1 activities of three water-soluble components were high indicated two parts of Arnebia euchroma showed better inhibitory activity against HepG2 cells; Arnebia euchroma significantly exhibited protective effect on mice with immune liver injury by inhibiting the release of inflammatory factors. The above work is only limited to the evaluation of anti-HBV, liver protection and anti-inflammatory activities, but its therapeutic material basis and mechanism of action remains unclear. Numerous studies have shown that NF-κB and MAPK signaling pathways are the main causes of HBx-mediated liver inflammation to hepatocellular carcinoma cells occurrence, development, rapid proliferation and metastasis. This project put forwards the research idea that "the effective part/effective components of Arnebia euchroma can exhibit anti-hepatoma effect by inhibiting the release of inflammatory cytokines and protein expression by inhibiting the activation of NF-κB and MAPK signaling pathways". This study will provide important evidence for elucidating the relationships among HBV infection, inflammation and hepatocellular carcinoma, and provide a theoretical basis for the development of Arnebia euchroma.

消化系统肿瘤被称为是“炎症相关性肿瘤”，从“炎癌转化"的角度研究炎症-肝癌发生发展的相关性及潜在机制意义重大。前期研究发现紫草乙醇及其热水提取物均具有较强的抗HBV活性，3个水溶性成分抗HBV及HIV-1活性显著；抗肿瘤筛选发现紫草2个部位对HepG2细胞均具有较好的抑制活性；紫草对小鼠免疫性肝损伤保护作用显著，抑制炎性介质的释放。上述工作基础仅局限于评价了紫草抗HBV作用和初步的保肝抗炎及抗肿瘤活性，但其药效物质基础及作用机制尚未明确。诸多研究表明，NF-κB 和MAPK信号通路是HBx介导肝脏炎症向肝癌细胞的发生、发展、快速增殖以及转移的主要原因。本课题提出“紫草有效部位/有效成分通过抑制NF-κB和MAPK信号通路的活化，从而抑制炎症因子释放和蛋白表达，发挥抗肝癌作用”的研究思路。该研究将为阐明HBV感染-炎症-肝癌相关性提供重要证据，同时为新疆软紫草开发提供理论依据。

紫草具有抗HBV和抗肿瘤活性，但其药效物质基础及作用机制仍有待进一步研究。本研究通过MTT、CCK-8法测定新疆软紫草石油醚部位、水提取物几种单体化合物抗炎、抗HBV、抗肝癌药效，初筛有效成分，并结合网络药理学筛选新疆紫草抗肝癌作用的药效物质群，预测核心成分和抗肝癌机制。研究建立了LPS诱导RAW264.7细胞炎症模型，并通过Griess和Western blot等方法开展体外抗炎初筛及机制探究。研究进一步通过H22肝癌荷瘤模型和DEN/CCl4慢性肝癌模型，并结合集落生成、细胞迁移侵袭和细胞凋亡等方法，探究异戊酰紫草素抗肝癌细胞恶性生物学行为作用。软紫草提取物包含6个脂溶性萘醌类和3个水溶性化合物。其中左旋紫草素通过抑制NF-κB信号通路p65和IKKα/β蛋白参与调控MAPK/NF-κB信号通路，从而发挥其抗炎作用。异戊酰紫草素通过调控PI3K/Akt和MAPK信号通路发挥抗肝癌作用。软紫草提取物制备工艺以及MAPK/NF-κB信号通路的确定为为紫草开发和临床应用提供了科学依据。