大黄蒽醌-半胱氨酸加合诱导的中药肝损伤机制研究

何首乌、大黄中大黄蒽醌的肝毒性问题是影响相关产品安全使用的风险因子，其致毒机制不清楚，是否致肝毒也存在争议，需要加强对其有可能产生的肝毒性及致毒机制研究。本课题首次发现大黄素可与肝脏内源性半胱氨酸形成加合物，而这种加合物呈现剂量和时间依赖性，与大黄素引起的肝细胞毒呈正相关。藉此提出大黄蒽醌肝损伤可能是由其引起半胱氨酸相关物质过度消耗及代谢异常，进而诱发肝毒性的原创性假说。本项目将应用体外人源性肝细胞模型和动物模型，给予不同剂量蒽醌单体、蒽醌混合物及含大黄蒽醌中药提取物，对不同时间点的生物样品进行蒽醌-半胱氨酸加合物检测、常规毒理指标检测、代谢组学和蛋白组学检测，筛选肝毒性差异标志物和差异代谢通路，阐述大黄蒽醌肝毒性机制。重点检测谷胱甘肽代谢通路、牛磺酸代谢通路异常及含半胱氨酸蛋白质的变性引起的功能异常及相关肝毒标志物，为含大黄蒽醌中药的毒性评价及安全、合理使用提供科学依据。

Anthraquinone-induced hepatotoxicity is the risk factor that affecting the security use of anthraquinone-containing herbs, such as rhubarb and radix polygoni multiflori. The mechanism of anthraquinone hepatotoxicity is still unclear and whether anthraquinones are the toxic factors or not is controversial. Therefore, researches on the hepatotoxicity of anthraquinone and the hepatotoxicity mechanism need to be further studied. We first discovered the anthraquinones-cysteine adduct in liver, which showed dose-and time-dependent along with emodin hepatotoxicity. Based on these findings, we proposed an original hypothesis that anthraquinone hepatotoxicity is probably induced by cysteine –related metabolic pathway disorder resulting from cysteine depletion. In this research, SD rats and human liver cells will be administered different doses of anthraquinone, anthraquinone mixture or anthraquinone-containing herb extracts. Anthraquinone –cysteine adduct, standard liver toxicology index, metabolic changes and proteome changes will be detected at different time points. Specially, cysteine-related metabolic pathway, such as glutathione synthesis, taurine metabolism and cysteine-containing protein metabolism, will be mainly detected. Based on these findings, we will systematically elaborated the mechanism of anthraquinone hepatotoxicity and discover specific anthraquinones hepatotoxicity biomarkers. This finding will provide scientific basis for toxicity evaluation and rational administration of anthraquinones containing drugs.