野马追的清热解毒药效物质基础研究

Traditional Chinese medicine scholars have long considered that the "heat clearing and detoxicating" effect and the "anti-pathogenic microorganism" effect equal, but that "equivalent mechanism" is difficult to fully elucidate its functions from the scientific point. We think heat clearing and detoxicating including "heat clearing" effect and "detoxicating" effect. We present the heat clearing and detoxicating drugs play " heat clearing" effect by antipyretic and anti-inflammatory,detoxicating effect through anti-pathogenic microorganisms. Based on the heat clearing and detoxicating drugs of anti-complement activity and control to heat syndrome characterized by the infectious diseases of the high degree of relevance, we propose the heat clearing and detoxicating drugs play the "heat clearing" effect by inhibiting complement activation to remove infectious diseases of fever and inflammation. Authentic and superior medicinal herbal of jiangsu Eupatorium lindleyanum DC. has anti-inflammatory, antibacterial and antiviral action. We performed the anti-complementary activity test shows that Eupatorium lindleyanum DC. has strong anti-complementary activity. We think Eupatorium lindleyanum DC.play the heat clearing effect by anti-complement. Based on the above experiments we will isolate and get the fractionations and compositions of "heat clearing" guided by anti-complement activity in vitro. We will validate the antipyretic effect of the heat clearing compositions by the endotoxin fever model in vivo, at the same time we will verify the anti-inflammatory effect of the heat clearing compositions by ELISA method. Based on the above research results we can elucidate the "heat clearing" effect and its pharmacodynamic materials of Eupatorium lindleyanum DC.. In addition we can elucidate the detoxification efficacy substance of Eupatorium lindleyanum DC. by the antibacterial and antiviral test. Both from theory or experiment we can all get the "heat clearing" and "detoxicating" pharmacodynamic materials of Eupatorium lindleyanum DC. based on traditional Chinese medicine theory. The research achievement of this project will provide the scientific basis for Eupatorium lindleyanum DC. quality control and the new ideas and research methods on heat clearing effect and its pharmacodynamic materials from the heat clearing and detoxicating drugs. At the same time for the "heat clearing" effect and the "detoxicating" effect of collaborative treatment of infectious diseases, which laid the foundation of scientific research.

中医药学者长期认为 "清热解毒"与"抗病原微生物"等同，但这一"等同的机制"难以全面阐明其作用的科学内涵，我们认为清热解毒包括"清热"和"解毒"功效，清热解毒药通过解热、抗炎发挥"清热"功效，通过抗病原微生物发挥"解毒"功效。基于清热解毒药抗补体活性与防治以热证为特点的感染性疾病的高度关联性，我们提出，清热解毒药通过抑制补体的过度激活来解除感染性疾病的发热和炎症，从而发挥其"清热"功效。江苏道地药材野马追有抗炎、抗菌抗病毒作用，我们进行的抗补体活性测试表明野马追有强抗补体活性，我们认为野马追通过抗补体而清热，在此基础上，本项目将体外抗补体活性导向分离清热成分，内毒素发热模型验证其体内解热作用，ELISA法验证其抗炎作用，从而阐明野马追的"清热"功效及药效物质；体外抗菌抗病毒测试，阐明野马追的解毒药效物质。本项目将为野马追的质量控制提供科学依据，并可能为清热解毒药的清热药效物质研究提供新思路

中医药学者长期认为清热解毒药通过“解毒”（抗病原微生物）而发挥作用，但这一“解毒”功效难以全面阐明其作用的科学内涵，依据中医理论，基于该类药有效防治重大感染性疾病及此类疾病与补体系统高度关联性，我们认为清热解毒包括“清热”和“解毒”功效，并且“清热”在该类药作用中扮演着重要的角色，清热解毒药通过抑制补体的过度激活来解除感染性疾病的发热和炎症，从而发挥其“清热”功效；通过抗病原微生物发挥“解毒”功效。江苏道地药材野马追具有抗炎、抗菌抗病毒作用，但药效物质不清楚，我们进行的抗补体活性测试表明其具有强抗补体活性，在此基础上，本项目采用UPLC/Q-TOF-MS法，分析野马追醇提取物的化学成分，野马追主要含有萜类和黄酮类化合物；体外抗补体活性测试，获得抗补体活性成分群为野马追倍半萜部位和黄酮部位，体外抗补体活性导向分离清热成分，分离鉴定了31个化合物，其中化合物6个新的倍半萜类化合物，3个新的二萜类化合物，12个化合物为首次从该植物中分离得到，化合物eupalinolide F ，eupalinolide I，eupalinolide K，木犀草素，异槲皮苷，槲皮素，芦丁，伪蒲公英甾醇等具有较强抗补体活性；脂多糖诱导大鼠炎性发热模型验证野马追的解热作用，野马追通过抑制肝组织中髓过氧化酶活性，降低了血清中肿瘤坏死因子-α、白介素-6含量而发挥解热作用。小鼠急性肺损伤模型及ELISA法验证了野马追的抗炎作用，野马追提取物，野马追黄酮部位、野马追水层及野马追倍半萜部位对脂多糖诱导的小鼠急性肺损伤具有保护作用，其作用机制与抑制炎症因子、降低补体C3水平和抗氧化有关。野马追的解毒与清热的药效物质并不尽相同，野马追黄酮和倍半萜类成分是野马追发挥“清热”功效的主要活性成分，二萜类化合物及部分黄酮类成分是野马追发挥“解毒”功效的主要活性成分；野马追乙醇提取物、野马追黄酮部位、野马追倍半萜部位、野马追70%乙醇水洗脱部位以及两种链状二萜化合物具有抗乙肝病毒活性；黄酮类化合物，木犀草素、泽兰叶黄素具有抗流感病毒活性；链状二萜化合物具有较强的抗菌活性；链状二萜化合物有弱的抗艾滋病毒活性。项目分别从“清热”和“解毒”功效，阐述了野马追的清热解毒作用及其药效物质; 课题完成了项目既定的研究内容，已发表论文SCI一篇，中文核心论文二篇，申报专利4项，达到项目既定的研究目标.