甘草及其主要活性成分甘草酸通过调节CYP2/CYP3A4缓解雷公藤多苷片所致肝损伤的作用机制研究

雷公藤多苷片（TGT）是中药雷公藤的代表制剂，临床用于治疗类风湿性关节炎（RA）疗效确切，但是其治疗窗口狭窄，易产生药源性肝损伤等副作用。“解毒圣药”甘草及其主要活性成分甘草酸具有确切的肝脏保护作用，与雷公藤配伍可达到减毒效果。围绕“甘草及甘草酸缓解TGT所致肝损伤的药效特点与作用机制”这一关键科学问题，根据申请人前期初步明确的CYP2和CYP3A4同时参与TGT所致肝损伤的发生以及甘草酸的肝脏保护效应机制，本项目拟通过结合细胞热转移分析（CETSA）与CRISPR/CAS9构建靶点敲低/敲除细胞模型的技术，进一步研究甘草及甘草酸调节CYP2和CYP3A4的上、下游靶标，全面阐明其通过调节CYP2/CYP3A4缓解TGT所致肝损伤的分子机制。综上，本项目的成功实施，将促进甘草及其主要活性成分甘草酸的解毒机制研究，指导TGT的临床合理应用，也为其他中药的作用机制研究提供方法学参考。

Tripterysium glycosides tablet (TGT), a representative preparation derived from Chinese herbal medicine Tripterygium wilfordii Hook (TWHF), has been effectively used in the treatment of rheumatoid arthritis. Due to the narrow treatment window, it has high risk to produce drug-induced liver injury and other side effects. Gancao (Glycyrrhizae radix et rhizome) and its major bio-active compound Glycyrrhizic acid (GA) have definite liver protective effect, which has been used to attenuate the side effect of TWHF. Preparatory experiments were preformed and demonstrated that CYP2 and CYP3A4 were found to be involved in both TGT-induced liver injury and the liver protection role of GA. To overcome TGT-induced liver injury, the key is to unravel the pharmacological actions and mechanisms of Gancao and GA. Built upon this, I will apply the state-of-art approaches cell thermal transfer assay (CETSA) and CRISPR/cas9 based target knockdown/ knockout system to comprehensively analyse the upstream and downstream targets of CYP2 and CYP3A4 in this project. Specifically, I aim to systematically reveal the pharmacological actions and molecular mechanisms of Gancao and its major bioactive compound GA in alleviating TGT-induced liver injury by regulating CYP2 / CYP3A4. In conclusion, this project aims to provide the foundation that can guide evidence-based usage of TGT in clinics and hold the promise of accelerating the research and development process of novel drugs derived from Gancao. In addition, it also provides us the opportunity and platform to research the pharmacological actions and mechanisms of other traditional Chinese medicines.