大黄蒽醌类是何首乌、大黄等中药引起肝肾损伤的主要致毒物。从蒽醌类自身代谢角度寻找解毒途径已成为关键科学问题，但目前其代谢调控机制与毒性关系尚不清楚。我们前期发现葡萄糖醛酸化是大黄蒽醌重要解毒途径，葡萄糖醛酸转移酶UGT1A9与UGT2B7为代谢关键酶，其表达极可能同时受转录因子HNF4A调控。为证实此代谢解毒调控通路，本项目拟：①采用人肝肾微粒体代谢模型，结合转录组、基因型数据及生物信息学分析阐明HNF4A-UGT1A9/2B7-大黄蒽醌代谢调控机制；②沉默/过表达细胞中HNF4A及UGT1A9/2B7，进行细胞毒性实验和代谢研究，阐明代谢调控与毒性关系；③以HNF4A-RNAi敲低小鼠及UGT酶抑制/诱导小鼠，结合肝肾毒性评价研究大黄蒽醌药代/毒代动力学，确证上述通路作用。研究结果可为发现UGT代谢酶诱导解毒剂促进大黄蒽醌类代谢降低其毒性提供科学理论依据，并为此类中药安全应用开辟新思路。

Polygonum multiflorum and Rheum officinale can lead to severe liver or kidney injury, and the Rheum Athraquinoines are the main toxic components. Clarifying the metabolic mechanism of these components is important to their detoxification. We have proven that glucuronidation is the main metabolic and detoxification pathway for Rheum Athraquinoines; UGT1A9 and UGT2B7 are the major metabolic enzymes; HNF4A may regulate the transcription of both UGT1A9 and UGT2B7 by binding their promoter region. In order to prove our hypothesis that the hepatic/renal detoxification of Rheum Athraquinoines is regulated by HNF4A-UGT1A9/UGT2B7-mediated metabolic pathway, we plan to: 1) Explore the individual glucuronidation of Rheum Athraquinoines in human liver and kidney microsomes, as well as the relationship between the UGT1A9/UGT2B7-mediated metabolism and the HNF4A expression based on the Genome-wide association study and RNA-seq association study; 2) Explore the detoxification of Rheum Athraquinoines through HNF4A-UGT1A9/UGT2B7-mediated pathway in vitro using HNF4A-overexpressing HepG2 cell, HNF4A-RNAi HepG2 cell, site-directed mutation cells, UGT1A9-overexpressing HEK293 and UGT2B7-overexpressing HEK293; 3) Explore the pharmacokinetics/toxicokinetics of Rheum Athraquinoines using UGT-inhibited mice, UGT-induced mice and HNF4A-RNAi FVB mice to further demonstrate our hypothesis. The study will clarify a new HNF4A-UGT1A9/UGT2B7-mediated regulatory metabolic pathway for the detoxification of Rheum Athraquinoines in liver/kidney and provide theoretical and experimental basis for the clinical security applications of traditional Chinese medicine such as Polygonum multiflorum and Rheum officinale.