下瘀血汤诱导肝星状细胞凋亡及逆转肝窦毛细血管化的抗肝纤维化药效物质基础研究

Xiayuxue decoction (XYXD) was first described in Jingui Yaolue, it has been used for the treatment of liver fibrosis in the clinical of modern traditional Chinese medicine. In our previous study, the ant-fibrotic effect of the components compatibility of emodin, amygdalin and the extracts of Eupolyphaga sinensis has been studied. It was found that the anti-fibrotic effect of XYXD was better than the components compatibility group. This result suggests that there may have more activity or synergistic effect chemical compositions (group) in XYXD. Basis on the anti-fibrotic pharmacological mechanism of XYXD through inhibiting hepatic stellate cells (Hepatic stellate cells, HSCs) activation, inducing HSCs apoptosis and reversing hepatic sinusoidal capillarization. In the project, the multidisciplinary theory and technology of traditional Chinese medicine, serum pharmacochemistry of traditional Chinese medicine, pharmacokinetics, chemistry of chinese materia medica and pharmacology were employed to study the serum pharmaceutical chemistry and pharmacokinetics of the main ingredient in different pathophysiological conditions after administration of XYXD. Studied on the effects of drug-containing serum by inhibited the activation of HSCs, induced apoptosis of HSCs and reversed the hepatic sinusoidal capillarization. The potential "active ingredients group" of anti-fibrotic in XYXD has been found and prepared. Then the effect and mechanism of anti-fibrotic of "active ingredients group" were performed in vivo by carbon tetrachloride (CCl4)-induced liver fibrosis of rats. The results were very important for clarification of the material basis of anti-fibrotic effect of XYXD and its new drug development.

下瘀血汤出自《金匮要略》，现代中医临床中常用于治疗肝纤维化。课题组前期将大黄素、苦杏仁苷与土鳖虫提取物组成组分配伍组进行抗肝纤维化研究，发现下瘀血汤全方组作用明显优于组分配伍组，提示下瘀血汤中可能存在活性更好或具有协同作用的化学成分（群）。基于下瘀血汤抑制肝星状细胞（Hepatic stellate cells，HSCs）活化、诱导HSCs凋亡与逆转肝窦毛细血管化发挥抗肝纤维化作用的药理作用特点，本课题运用中医药学、中药血清药物化学、药代动力学、中药化学、药理学等学科的理论与技术，研究下瘀血汤在不同病理生理状态下的血清药物化学与药代动力学，评价含药血清抑制HSCs活化、诱导HSCs凋亡及逆转肝窦毛细血管化作用，揭示其抗肝纤维化的潜在“活性成分群”，制备“活性成分群”组合物并进行抗肝纤维化作用验证与机制探讨。研究结果对于阐明下瘀血汤中抗肝纤维化作用的药效物质基础与其新药开发等具有重要意义。

下瘀血汤出自《金匮要略》，现代中医临床中常用于治疗肝纤维化。但药效活性成分并不清楚，严重制约了其新药开发与临床运用。本课题基于下瘀血汤抑制肝星状细胞（HSCs）活化等药理作用特点，运用中医药学、中药血清药物化学、药代动力学、中药化学、药理学等学科的理论与技术开展了系列研究。鉴定了下瘀血汤中115种主要化学成分，完成下瘀血汤在不同病理生理状态下13种主要成分大鼠体内的药代动力学研究；基于体内外药效研究，明确乙酸乙酯萃取部位是其主要活性部位，抑制HSCs活化、增殖及诱导细胞周期阻滞，选择性激活JNK和p38 MAPKs信号通路调控线粒体凋亡途径而诱导HSCs凋亡，抑制肝窦内皮细胞（LSECs）失窗孔化等是乙酸乙酯部位抗肝纤维化主要作用机制；完成了对乙酸乙酯部位19种主要化学成分体外活性筛选，并结合含药血清谱效关系研究，明确Rhe、Amy等是下瘀血汤抗肝纤维化的主要活性成分；并创新性发现下瘀血汤对非酒精性脂肪性肝炎的肝纤维化具有良好的干预作用，乙酸乙酯部位同样也是其主要活性部位，Rhe与Amy是其主要活性成分，并采用均匀设计筛选出100 mg/kg Rhe与0.25 mg/kg Amy为最佳配伍组合。本研究明晰了下瘀血汤抗肝纤维化的主要药效活性成分，可为其中药成分组合新药开发及下瘀血汤临床运用提供部分参考。