基于肝窦内皮细胞与星状细胞关系探讨当归补血汤组分复方抗肝血管新生与纤维化的作用机制及配伍特点

The former Natural Science Foundation indicated that the Dangdui Buxue formula (DBF) inhibited liver fibrosis and angiogenesis in CCl4 rats. The related active components were found and formed a new formula. Recent researches confirmed that angiogenesis is the key pathologic mechanism of liver fibrosis. The injury and activition of liver sinusoidal endothelial cells (LSECs) and hepatic satellite cells (HSCs) is the cytology foundation of angiogenesis. In this project, we choose the perspective of LSECs injury causing the activation of HSCs. The fibrotic animal models, 2 cell co-cultural systems and zebrafish angiogenesis models will be used. By following the signal pathways related to angiogenesis like "Oxidative injury / NF-κB", "VEGF or PDGF/MAPK", the key chemical mechanism of anti-angiogenesis and anti-fibrosis of DBF components formula will be observed. Secondly, the orthogonal design will to be used to clear the compatibility characteristics of DBF components formula in vivo and in vitro based on the major targets of angiogenesis. This project will confirm the hypothesis of "components formula inhibit angiogenesis and liver fibrosis by ways of protecting the LSECs from injury and regulate the relationship of LSECs and HSCs with the prior effect to DBF. There's a compatibility characteristics on the level of molecular pharmacology." This study will promote the development of anti-fibrosis therapy by using traditional Chinese medicine, and lay the foundation for discovering new "Chinese herbs".

已结题青年基金工作表明:当归补血汤可通过抑制肝血管新生而抗肝纤维化，具有相应活性成分，并形成新的组分复方。新近研究认为肝血管新生是肝纤维化关键病理机制，其细胞学基础在于肝窦内皮细胞与星状细胞的损伤活化及其相互影响。本项目首先从肝窦内皮细胞损伤导致星状细胞活化的角度，围绕"氧化损伤/NF-κB"、"VEGF或PDGF/MAPK"等血管新生信号通路，通过肝纤维化动物模型、2种细胞共培养体外模型、斑马鱼等模式生物，观察"当归补血汤组分复方"抗肝纤维化血管新生的关键药理机制；然后针对血管新生的主要靶标，采用正交设计，在分子水平明确该复方的组分配伍特点。以验证"当归补血汤组分复方通过保护肝窦内皮细胞损伤，并调节该与肝星状细胞关系，从而抑制血管新生与抗肝纤维化；组分复方作用优于原方，且在分子水平具有配伍特征"的科学假说。以促进中医药抗肝纤维化优势学科的发展，并为组分复方创新中药奠定坚实基础。

前期工作表明:当归补血汤可通过抑制肝血管新生而抗肝纤维化，具有相应活性成分，并形成新的组分复方。新近研究认为肝血管新生是肝纤维化关键病理机制，其细胞学基础在于肝窦内皮细胞与星状细胞的损伤活化及其相互影响。本项目围绕“氧化损伤/NF-κB”等血管新生信号通路，通过2 种肝纤维化动物模型、2种细胞体外模型、斑马鱼等模式生物，观察“当归补血汤组分复方”抗肝纤维化血管新生的关键药理机制；然后针对血管新生的主要靶标，采用正交设计，在分子水平明确该复方的组分配伍特点。研究结果表明，组分复方可减轻DMN、CCl4模型鼠肝组织胶原沉积和星状细胞活化，下调肝组织VEGF、PDGF等表达，抑制肝窦内皮细胞的增殖、迁移及管腔形成，抑制肝血管新生。其作用机制与调控PARP-1/NF-κB信号通路保护肝窦内皮细胞损伤、抑制其过度增殖从而减少肝星状细胞活化有关。黄芪来源的毛蕊异黄酮保护肝窦内皮细胞损伤影响最大，当归来源的欧当归内酯A抑制内皮细胞增殖影响最大，体现了组分复方在分子水平的配伍特征。对其中主要成分欧当归内酯A的体内外实验发现，该成分可有效抑制纤维化血管新生。项目实施过程所形成和完善的肝纤维化血管病理评价模式有助于促进中医药抗肝纤维化创新药物研发，并为组分复方配伍理论研究奠定实验基础。