苗药黑骨藤在不同生理状态下的药动学及代谢特征研究

Periploca Forrestii, one of the ten key Miao’s medicinal plans in Guizhou province, is widely used for the treatment of rheumatic joint pain and traumatic injury in Guizhou ethnic minority regions. The investigation of our research team has proved for the first time that the active water-soluble ingredients in Periploca Forrestii had outstanding curative effect on rheumatoid arthritis (RA) while the material base for efficacy, action mechanism and process in vivo still remain unclear. The studies are planned to be conducted focusing on the effective ingredients of Periploca Forrestii . The following research work will be involved: (1) to conduct the study on the transitional constituents and fingerprint pharmacodynamics between the control group and animal pathological model group, then to treat the data and manage to the find out the relationship between the characteristic constituents and the anti-inflammatory activity and to identify the PK markers. (2) to build up the PK-PD models in rats and to discover the PK-PD relation under physiological and pathological conditions in order to reveal the law of anti-inflammatory action mechanism and the dynamic process in vivo of effective ingredients in Periploca Forrestii. (3) to study the effects on the metabolism network of model rats by metabolomics approach so as to discover the regulation mechanism of related endogenesis substance and metabolism path. Above research results will provide the evidence for the effective ingredients and mechanism of action of Periploca Forrestii.

黑骨藤在贵州苗族地区广泛以内服外用治疗风湿关节痛、跌扑损伤，为我省十大苗药之一。课题组首次研究发现黑骨藤水溶性有效组分治疗类风湿性关节炎疗效独特，但其药效物质基础、作用机制及体内过程尚不明确。本研究旨在以黑骨藤有效组分为研究对象，（1）开展黑骨藤在正常大鼠和病理模型大鼠的体内移行成分及谱效关系研究，分析其特征差异以及与抗炎活性的相关性；（2）建立黑骨藤PK-PD模型，分析正常和病理状态下药代动力学与药效动力学的相关性，揭示其抗炎作用的效应成分及体内动态变化规律；（3）利用代谢组学方法，探讨黑骨藤对炎症动物模型代谢网络的影响，确定其对机体内源性代谢物及代谢通路的调控机制。上述研究结果将为阐明黑骨藤的药效物质基础和作用机制提供科学依据。

黑骨藤在贵州苗族地区广泛用于治疗风湿关节痛、跌扑损伤，为我省十大苗药之一。课题组前期研究发现黑骨藤提取物治疗类风湿性关节炎（RA）疗效独特，但其药效物质基础、作用机制及体内过程尚不明确。本项目完成了基于AA大鼠和MH7A细胞模型的黑骨藤抗RA作用研究，可明显降低AA大鼠TNF-α、IL-1β和RF的含量，其作用机制与抑制NF-κB通路及MAPK通路有关。完成了黑骨藤提取物在正常大鼠和AA大鼠体内的移行成分研究，通过抗炎效应成分谱效关系分析确定了PK markers。通过比较黑骨藤提取物在正常和AA大鼠血浆、尿液和粪便中的代谢产物，发现在正常和模型大鼠体内均发生较广泛的I相和II 相代谢。开展了正常和AA大鼠口服和静注给予黑骨藤提取物后主要活性成分的药动学特征研究，发现黑骨藤提取物在AA大鼠体内代谢快、排泄快，分析其差异原因为病理状态下药物代谢酶谱可能会发生改变，从而影响药物代谢、分布与消除。研究建立了黑骨藤的PK-PD模型，提示药效指标IL-1β、RF、TNF-α与黑骨藤3种活性成分的浓度存在良好相关性。采用UPLC-MS/MS及GC-MS技术开展了基于AA大鼠模型的代谢组学研究，结合代谢通路分析表明类风湿性关节炎可能与体内的亚油酸、甘油磷脂、花生四烯酸、鞘脂、氨基酸、脂肪酸、糖类等代谢通路紊乱有关，黑骨藤治疗后可使紊乱的代谢通路恢复正常水平而产生抗RA作用。建立了基于黑骨藤药效物质的多指标质量控制新方法。上述研究为黑骨藤创新药物研制及药材资源深度开发提供了科学依据。通过项目实施，发表研究论文13篇，申请发明专利1项，研究成果荣获了贵州省科技进步一等奖。