基于“玄府-络病-风药”理论研究桂枝茯苓丸经方新用防治肝纤维化的作用及机制

In TCM, xuán fǔ and collaterals in the liver are the gateway and place for the exchange of microscopic substances and information, and the main pathogenesis of hepatic fibrosis (HF) is the accumulation of dampness toxin and blood stasis caused by invasion of dampness toxin, qi deficiency and blood stasis block liver collaterals. In that way, HF can be classified into the category of collateral disease. In western medicine, the hepatic sieve formed by windows of hepatic sinusoid endothelial cells is the microchannel for substance exchange. One of the key pathological mechanisms of HF is Hepatic microcirculatory disturbance, and the hepatic stellate cells (HSCs) activation and hepatic sinusoid capillarization are the central links. To sum up, TCM and western medicine share the same connotation in understanding the physiological structures of liver and pathogenesis of HF. Based on the thought of treating different diseases with the same method, Guì Zhī Fú Líng Wán (Cinnamon Twig and Poria Pill), functioning on dissolving stasis to dissipate concretions, is corresponding to the pathogenesis of stagnation of liver collaterals. According to the theory of ‘xuán fǔ-luò bìng-fēng yào’, Guì Zhī Fú Líng Wán, manifesting characteristics of fēng yào (wind medicinals), could open xuán fǔ in the liver and unblock liver collaterals to maintain the liver’s nature of preferring free activity.To this end, by applying methods of molecular biology, immunology and proteomics, the study intends to verify the effectiveness of Guì Zhī Fú Líng Wán in the preventing and treating HF and to investigate its effect on the activation of HSCs and the capillarization of hepatic sinusoid. In the end, experimental evidences are provided to support the understanding of stagnation of liver collaterals in HF and the application of Guì Zhī Fú Líng Wán in prevention and treatment of HF.

肝内玄府与肝络具有微观性物质交换与信息交流的通道性，肝纤维化主要病机变化为湿毒内蕴、气虚血瘀，导致湿毒瘀互结，入肝滞络，可归属中医“络病”范畴。此与肝窦内皮细胞窗口构成的肝筛是窦周间隙内外物质交换的微观通道，肝脏微循环障碍是肝纤维化关键病理机制之一，肝星状细胞激活与肝窦毛细血管化是肝脏微循环障碍的中心环节的生理结构认识与病机变化认识内涵一致。基于中医异病同治之思想，具有化瘀消癥功效的桂枝茯苓丸与肝纤维化肝络瘀阻之关键病机呈“方证对应”。基于“玄府-络病-风药”理论，桂枝茯苓丸具风药之性，与风气之脏同气相求，能开肝内玄府、畅肝络，持肝喜条达之秉性。由此，本研究拟通过对CCl4肝纤维化模型大鼠的治法方药干预，运用分子生物学、免疫学及蛋白组学等方法验证桂枝茯苓丸防治肝纤维化的有效性，考察其对肝星状细胞激活与肝窦毛细血管化的影响，寻找“肝络瘀阻”病机理论的实证依据，并为经方新用提供一定的实验依据。

肝纤维化（hepatic fibrosis, HF）是全球公共卫生组织关注的主要问题之一。目前，针对单一靶标研发的化学药物或生物药物对复杂病理机制的HF难以取得良效。越来越多的研究证实了中医药防治HF具有独特优势。中药复方具有整体性、多组分和多靶点等特点，对病理机制复杂的疾病，可整体调节多个环节，通过多靶点产生协同效应，从而取得良效。中医学肝内玄府与肝络具有微观性物质交换与信息交流的通道性，肝纤维化主要病机变化为湿毒内蕴、气虚血瘀，导致湿毒瘀互结，入肝滞络，可归属中医“络病”范畴。此与西医学肝窦内皮细胞窗口构成的肝筛是窦周间隙内外物质交换的微观通道，肝脏微循环障碍是肝纤维化关键病理机制之一，肝星状细胞激活与肝窦毛细血管化是肝脏微循环障碍的中心环节的生理结构认识与病机变化认识内涵一致。基于中医异病同治之思想，具有化瘀消癥功效的桂枝茯苓丸与肝纤维化肝络瘀阻之关键病机呈“方证对应”。本研究以CCl4肝纤维化大鼠、HSC-T6细胞为研究对象，研究了桂枝茯苓丸防治肝纤维化的作用及机制。结果提示桂枝茯苓丸可改善CCl4肝纤维化大鼠肝脏组织病理形态改变，胶原沉积及肝功能损伤；抑制HSC-T6细胞的活化与增殖，改善ECM异常生成。其可能是通过调节TGF-β1/Smad2信号通路调控HSCs，及通过调节VEGF/VEGFR-2信号通路阻止病理性血管增生和调节P62/Keap1/Nrf2信号通路抑制氧化应激反应，从而调控肝窦毛细血管化以发挥防治肝纤维化的作用。