基于胶原型关节炎大鼠模型比较蛋白质组学的维药槌果藤作用机制及靶点研究

Rheumatoid arthritis was an autoimmune diseases characterized by chronic synovial inflammation and joint destruction, which was also one of the common and the disabled joint disease in the department of rheumatology. Most traditional medicine in the rheumatoid arthritis’s clinical treatment had side effects, and emerging biological agents had some shortcomings such as short half-life, expensive and so on. So it seemed particularly urgent to look for safe, effective and cheap drugs. Capparis Spinosal, an Uygur medicine, was a special medical resources in Xinjiang region, which was oftern used to treat rheumatoid arthritis and gout by people. Our previous animal experiments and clinical studies both had shown Capparis Spinosal had good effective for Rheumatoid arthritis. This subject was designed to extract effective components of Capparis Spinosal, authenticate the active ingredients of Capparis Spinosal in vitro/in vivo by UPLC -PDA ; screen the different proteins of the type Ⅱ collagen induced arthritis rat model which was dealed with Capparis Spinosal’s active ingredients before and after by the iTRAQ technology, bioinformatics analysis was used to assess the different proteins screened from the pathway, function and protein network, which was in order to find the possible drug targets of Capparis Spinosal against Rheumatoid arthritis, provide the basis for Recombinant protein drugs’s research in future. Research on the mechanism of the rich uygur medical resources in Xinjiang also further develop the characteristics of medical resources in Xinjiang.

类风湿关节炎是一种以慢性滑膜炎症、关节破坏为特征的自身免疫性疾病，也是风湿科常见和致残的关节病之一。临床治疗类风湿关节炎的传统药物多有副作用，而新兴生物制剂又存在半衰期短、价格昂贵等缺点。故寻找安全、有效、廉价的药物显得尤为迫切。维药槌果藤是新疆特色医药资源，民间验方常用于治疗风湿性关节炎、痛风等。课题组前期动物实验及临床研究均显示其具有较好抗类风湿性关节炎作用。本课题旨在提取维药槌果藤有效成分，利用UPLC-PDA法体内/体外鉴定其药效活性成分；利用比较蛋白质组学iTRAQ技术对槌果藤活性成分干预胶原型关节炎大鼠模型前后差异蛋白进行筛选，从经典通路、功能分析以及通路网络三个方面对筛选出的差异蛋白进行生物信息学分析，以期找到维药槌果藤抗类风湿性关节炎可能的药物作用靶点，为今后蛋白质重组药物的研发奠定基础。对新疆丰富维吾尔医药资源的机制研究，也利于进一步开发新疆特色医药资源。

风湿关节炎是一种以慢性滑膜炎症、关节破坏为特征的自身免疫性疾病，也是风湿科常见和致残的关节病之一。维药槌果藤是新疆特色医药资源，民间验方常用于治疗风湿性关节炎、痛风等。课题组前期动物实验及临床研究均显示其具有较好抗类风湿性关节炎作用。本项目成功分离出维药槌果藤的各种药物组分，通过UPLC-MS/MS，建立了槌果藤不同提取物的药物指纹图谱，为今后继续探索研究新型化合物创造了条件。在体外实验中发现槌果藤石油醚提取物、氯仿提取物与乙酸乙酯提取物均具有较好的抗炎效果，其中乙酸乙酯提取物显著抑制了LPS诱导的RAW264.7巨噬细胞的NF-κB P65蛋白的核易位。进一步研究发现维药槌果藤活性成分阿魏酸、原儿茶醛、芥子酸均能发挥抗炎作用。LPS介导的NF-κB p65核转位被阿魏酸明显阻断，推测阿魏酸可能是维药槌果藤抗炎的活性成分之一。利用比较蛋白质组学iTRAQ技术对槌果藤活性成分阿魏酸干预胶原型关节炎大鼠模型前后差异蛋白进行筛选，发现其与维药槌果藤组的差异蛋白具有相似性，这些差异蛋白主要分布在代谢领域、细胞粘附、细胞周期和固有免疫信号通路领域。