基于循环exosomes的川楝子肝毒性生物标志物发现及毒性机制研究

药物性肝损伤是中药的主要不良反应，但当前由于缺乏有效的中药肝毒性评价方法，难以对中药肝毒性的物质基础及毒性机制进行准确识别与研究，阻碍了中药现代化与国际化的发展。本项目基于exosomes有望为药物肝毒性评价领域带来重大突破的前景，提出一种利用循环exosom-es内部和外部miRNAs的分子模式来识别中药肝毒性机制及发现毒性生物标志物的全新方法，从中药“多组分复杂体系”的特点出发，着重研究川楝子组分/单一成分在作用时间及剂量依赖下的小鼠肝毒性表型及机制的异同，系统分析血清exosomes内部miRNAs分子的表达谱及exos-omes外部肝特异性miRNAs的表达水平，筛选川楝子肝毒性发生、发展过程中关键的分子事件，揭示毒性机制，对exosomes内部特征性表达的miRNAs进行细胞水平跨种属的验证研究，寻找发现潜在的肝毒性生物标志物，建立基于循环exosomes的中药肝毒性评价新方法。

Drug-induced liver injury has been the most frequently adverse reaction of traditional Chinese medicine. Nevertheless, it is difficult to accurately identify toxic components and mechanisms of traditional Chinese medicine-induced hepatotoxicity as there is a lack of effective screening and evaluation methods, which is the key bottleneck of modernization and internationalization of traditional Chinese medicine. As exosomes are expected to bring a major breakthrough in the field of drug-induced hepatotoxicity, this project first proposes that miRNAs distribution patterns inside or outside of circulating exosomes may reflect the mechanisms of traditional Chinese medicine-induced hepatotoxicity and identify new biomarkers. As traditional Chinese medicine is a multi-component complex system, this project will investigate the dose- and time-dependent phenotypes and mechanisms of the components or single component of fructus toosendan-induced hepatotoxicity. The expression levels of miRNAs inside or liver-specific miRNAs outside of circulating exosomes of each treatment will be detected. The key molecular events and the mechanisms of fructus toosendan-induced hepatotoxicity will be determined. In order to identify cross-species biomarkers, exosomes-based potential biomarkers will be verified on cellular level. This project will establish a new method to evaluate traditional Chinese medicine-induced hepatotoxicity based on circulating exosomes.