基于代谢组学雷公藤治疗强直性脊柱炎药效特点和作用机理研究

Ankylosing spondylitis (AS) is one of major disease that damages human health severely. Clinical studies have identified that Tripterygium Wilfordii plays an important role in AS treatment, but the exact mechanism has not been clarified. Considering that metabonomics has great potential for elucidating the mechanism and efficacy of traditional Chinese medicine, as well as the pathogenesis of complex diseases, in the current project, an RP/HILIC-UPLC-TOFMS metabonomic approach is employed to reveal the evolution process of rat AS disease, demonstrate and compare the action mechanism and therapy properties of Tripterygium Wilfordii on the prevention and treatment of rat AS. The biomarkers related to AS pathogenesis and therapeutic effects of Tripterygium Wilfordii would be screened out and identified, and their related metabolic pathways would be speculated, which could deepen the understanding of the pathogenesis of AS, facilitate early clinical diagnosis and treatment on AS, promote the rational development and application of Tripterygium Wilfordii, and intensify the scientific understanding on the connotation of the traditional Chinese medicine theory on AS therapy.

强直性脊柱炎(AS)是严重危害人类健康的疾病，临床研究已证实雷公藤在治疗AS中发挥至关重要的作用，其确切机制尚未阐明。而代谢组学则在阐明中药药效特点和作用机理以及复杂疾病发病机理方面极具应用潜力。基于此，本项目拟采用RP/HILIC-UPLC-TOFMS代谢组学方法，在AS模型大鼠水平结合临床样本，从内源性代谢产物水平揭示AS疾病发生发展的演变过程，确定疾病进程相关的生物标志物和生理生化代谢通路，深化对AS发病机理的认识，指导临床AS早期诊断和治疗；从代谢轮廓和生物标记物水平科学考察雷公藤对AS的防治作用，初步阐明其药效特点和作用机理，以指导雷公藤的合理开发和应用，深化对中医药抗AS理论内涵的科学认知。

背景：强直性脊柱炎(AS)是严重危害人类健康的疾病，临床研究已证实雷公藤在治疗AS中发挥至关重要的作用，其确切机制尚未阐明。基于此本项目拟采用多组学联合方法，在AS模型小鼠水平结合临床样本，从内源性代谢产物水平揭示AS疾病发生发展的演变过程，确定疾病进程相关的生物标志物和生理生化代谢通路，深化对AS发病机理的认识，指导临床AS早期诊断和治疗；以指导雷公藤的合理开发和应用，深化对中医药抗AS理论内涵的科学认知。.方法：构建了强直性脊柱炎小鼠模型，分别给予抗炎药西乐葆和雷公藤红素灌胃治疗；借助多组学方法观察AS小鼠的血清代谢产物变化规律，以期望发现雷公藤红素高剂量治疗AS诊断及病情进展的生物标志物；前期在临床上收集了15例AS患者血浆样本采用TMT技术以及数据库进行信息分析构建网络图，并采用血清蛋白组学技术寻找一系列疾病标志物来联合诊断AS；又借助UPLC-QE分析140例AS患者与健康人群的血清样本，确定脂质诊断标志物；开展了运用温肾蠲痹方治疗强直性脊柱炎肾虚督寒证的临床疗效评价临床研究。.结果：体内实验显示雷公藤红素剂量依赖性改善模型鼠脊柱变形和椎间盘骨质增生现象；代谢组学结果显示苹果酸，苯丙氨酸等代谢水平在AS模型组中下调，雷公藤红素高剂量给药后其代谢水平上调，甘氨胆酸等胆汁酸代谢水平在AS模型组中上调，雷公藤红素高剂量给药后这些代谢物呈现下降趋势；蛋白组学发现CRP、SAA1与ORM2三种蛋白联合对AS的诊断具有高准确性和灵敏性，有望开发为强直性脊柱炎的标志物；脂质组学显示DAG18:1-20:4等脂质有助于AS的诊断。.结论：本研究从多组学角度，研究AS患者血清中的内源性代谢产物，进而全面揭示AS的发病机制以及寻找理想的用于AS诊断的标志物。观察了雷公藤治疗AS的作用效果，为初步阐明其药效特点和作用机理奠定基础，并指导雷公藤的合理开发和应用。