维药天山堇菜和一枝蒿治疗重症流感的物质基础研究

The Xinjiang is located in the Eurasian continent that a swine influenza (flu) virus is typically found, and is regarded as one of the major area for bird flu outbreak due to bird migration. The traditional Uighur medicines (TUM) have its own theory and clinical experiences for fighting the flu pandemic, especially for the severe flu. But the active substances and mechanisms of actions have not yet been elucidated. We propose a new "heat-clearing" conception that consists of complement inhibition, antipyretic action and anti-inflammation, based on which the TUM might be scientific explained, along with considering the known "detoxifying" (anti-virus and antibacterial) conception. The new idea of this proposal is to establish an integration method of in vitro "heat-clearing" (anti-complement) and "detoxifying" (anti-virus) screening and in vivo evaluation (antipyretics and actions on influenza A-induced acute lung injury) for revealing the potential active components and mechanisms of action of the TUM, Viola tianshanica and Artemisia rupestris. The achievements of this research will provide a new insight and idea for study on the active substances and mechanisms of severe flu of TUM.

新疆地处欧亚大陆这一世界甲流爆发典型区域，同时作为我国三大候鸟迁徙地之一也是禽流感的主要爆发地之一。维药作为我国四大民族药物之一，在防治流感特别是以急性肺损伤为特征的重症流感上有着较强的民族特色，具有独特的理论和实践经验。但维医药治疗该类疾病的物质基础和科学内涵尚未得知。在前人大量探索性研究和我们前期研究的基础上，并结合中医药和现代医药学的方法和技术，我们提出以"抗补体-解热-抗炎"三位一体新的"清热"概念，结合"抗流感病毒抗肺炎球菌"的"解毒"概念，全面揭示特色清热解毒维药天山堇菜和一枝蒿治疗重症流感的物质基础。本项目力图构建体外"清热"(抗补体)和体外"解毒"(抗病毒抗菌)筛选模型与体内药效(解热、甲型流感病毒诱导的急性肺损伤)验证一体化筛选和评价方法，由点到面，由外及内全面阐述维药在治疗该类疾病方面的药效物质基础和科学内涵。该研究将为维药防治重症流感的药效物质基础提供新的思路和方法。

本课题通过体外“清热”(抗补体)和体外“解毒”(抗流感病毒抗菌)筛选模型与体内药效(LPS诱导的急性肺损伤)评价相结合阐明了维药天山堇菜和一枝蒿等治疗流感的药效物质。重点探讨了天山堇菜基于抗补体活性和抗菌活性筛选出的清热和解毒活性部位的化学成分，从中分离鉴定了34个化合物。采用体外溶血法对这些单体化合物进行了抗补体活性经典途径和旁路途径的活性评价，并对部分活性化合物进行了抗补体活性作用靶点研究。在此基础上，采用体内LPS诱导的小鼠急性肺损伤模型，对活性石油醚部位和乙酸乙酯部位的治疗作用进行了评价，发现其醇提物的石油醚部位对小鼠急性肺损伤有良好的治疗作用。为了更加全面地阐明天山堇菜的药效物质基础，我们还首次建立了UPLC-ESI-qTOF-MS/MS法表征了其解毒部位的31个黄酮苷类成分，以及首次建立了针捕集(needle trap)-GC-MS法全面表征了其清热部位的104个挥发性成分。类似的，我们还对天山堇菜类同品甜地丁和另一个维药一枝蒿进行了清热与解毒的药效物质基础研究。该研究将为维药防治重症流感的药效物质基础提供新的思路和方法。