基于当归四逆汤配伍及显效状态的含马兜铃酸中药细辛效毒变化规律研究

Asarum belongs to aristolochic acids (AAs)-containing traditional Chinese medicine. Danggui Sini Decoction (DSD) is the prescription of Asarum as the main drug, which has remarkable therapeutic effect of dysmenorrhea. However, due to the long administration period and the need of regular cyclic administration, its safety has attracted much attention. Our previous studies have found that the same traditional Chinese medicine expresses different in vivo components in different compatible environments in order to realize the effect expression of compatibility and attenuation effects. Therefore, we hypothesize that the concentration of AAs in Asarum was decreased or not expressed under the condition of prescription compatibility and treatment, so as to achieve the effect of reducing toxicity and at the same time exerting the effect of Asarum assisting warm the channels and free the vessels of DSD. Thus, we will adopt metabolomics technology to characterize AAs and Asarum toxicity biomarkers, and dysmenorrhea model biomarkers firstly. Then, the biomarkers of dysmenorrhea model and toxicity were used as references, to reveal the effects, mechanism and in vivo active ingredients of DSD on dysmenorrhea animal model, as well as targeting characterization and analysis of toxicity biomarkers and changes of AAs in vivo during the administration cycle. We prospect to bring it to light that the changing law of effectiveness and toxicity, as well as attenuation mechanism of AAs-containing traditional Chinese medicine of Asarum based on the compatibility and effective state of DSD, to provide scientific basis for ensuring the safety and effectiveness of clinical medication of Asarum and Chinese patent medicines containing Asarum.

细辛属于含马兜铃酸（AAs）类中药，当归四逆汤是细辛作为主要药物的方剂，治疗痛经确有疗效，但由于给药周期较长且需定期循环给药，其安全性备受关注。课题组前期研究发现，同一中药在不同配伍环境表达不同的体内成分，从而实现配伍的效应表达及减毒作用。所以我们推测，在当归四逆汤配伍及治疗状态下，其他药物使细辛中AAs血中浓度减低或不表达，从而实现减毒作用同时又发挥了细辛佐使当归四逆汤温经通脉之效。由此，本课题采用代谢组学技术，首先表征AAs、细辛毒性标志物及痛经模型标志物；在显效状态下，以痛经模型标志物及毒性标志物为参数，评价当归四逆汤干预痛经动物模型效应、效应机制、体内显效成分，靶向表征及分析给药周期内毒性标志物及体内AAs变化规律，揭示当归四逆汤配伍环境及显效状态下含AAs中药细辛的效毒变化规律及减毒机制，为保障细辛及含细辛中成药临床用药安全与有效提供科学依据。

中药安全性一直是中药国际化进程中重点关注的问题，其中，马兜铃酸引起的肾毒性事件不仅涉及药材品种众多，更涉及众多确有疗效中成药的使用问题。细辛是含马兜铃酸的中药，是国内乃至国际需求量较大的常用中药，然而，中医临床给药既不是使用单一化学成分，也不是使用单一中药材，通常是以方剂的形式给药。基于此，本研究以原发性痛经大鼠为研究对象，采用中医方证代谢组学研究策略，在当归四逆汤显效状态下探讨含马兜铃酸中药细辛的效毒变化规律。研究发现，当归四逆汤可显著回调与原发性痛经相关的临床生化指标，并且对于原发性痛经相关的代谢生物标志物有不同程度的回调作用，涉及亚油酸代谢、甘油磷脂代谢、花生四烯酸代谢等；表征了当归四逆汤体外化学成分150个，采用PCMS关联分析确定槲皮苷、阿魏酸、绿原酸、甘草苷、芍药苷等11个化合物为当归四逆汤治疗原发性痛经的潜在药效物质基础；表征了基于原发性痛经的马兜铃酸潜在血液毒性生物标志物7个，潜在尿液毒性生物标志物19个，涉及固醇激素生物合成、甘油磷脂代谢，色氨酸代谢等与代谢通路，上述标志物以及代谢通路可以提前预测毒性反应的发生及进展；此外，从成分、代谢轮廓、代谢通路以及药效/毒效生物标志物为基础对基于当归四逆汤配伍及显效状态的含马兜铃酸中药细辛效毒变化规律进行阐述，表明了当归四逆汤通过调节多环节的潜在毒性生物标志物，进而起到配伍减毒的作用，并且当归四逆汤对潜在毒性生物标志物的调节作用强于缺细辛当归四逆汤，表明了细辛在当归四逆汤方剂中的重要性。本研究为细辛及细辛中成药的临床用药安全和有效提供了科学依据及参考。