基于氨基酸组学寻找与确认药物性肝损伤早期发现及疗效评价的生物标志物

The widely used of drug combination and new drugs increase the risk of drug-induced liver injury. Deficiency of specific biomarkers is the bottleneck of early discovery and efficacy evaluation of drug hepatotoxicity. We had completed the investigation of amino acid metabolic profiles in rats related to hydrazine drug induced hepatotoxicity. The results of this preliminary study provide ideas and basis for continued and further metabonomics research in clinic. To obtain specific biomarkers of drug-induced liver injury for early discovery and efficacy evaluation, an integrated approach of rapid resolution liquid chromatography mass spectrometry (RRLC-MS) in positive and negative ionization combining with multivariate data analysis is developed for metabonomic analysis on human serum and urine samples. Through time-course changes in potential biomarkers detected using a metabonomic approach and their relativity in metabolic pathway, biological significance of amino acid biomarkers was confirmed. In addition, the standardized metabonomic analysis method is established by controllable parameters assessed by biological control samples and mixed standard solution. This study will provide important basis for diagnosis and treatment of drug induced liver injury in clinic.

联合用药与新药的广泛使用加大了药物性肝损伤发病风险，特异性标志物的缺乏成为药物性肝损伤早期发现和预后判断的瓶颈问题。课题组前期已完成肼类药物肝损伤的氨基酸组学特征及其作为肝损伤早期预警标志的可行性研究，已获研究结果为延续并深入至临床药物性肝损伤的代谢组学研究提供思路与基础。本项目拟采用正负离子检测模式相结合的RRLC-MS/MS 技术结合多变量统计分析方法，针对健康者、药物性肝损伤初诊（单纯抗菌药物与抗肿瘤药物引发的肝损伤）及不同治疗阶段的血浆和尿液样本开展组学研究，旨在寻找及确认与药物性肝损伤早期诊断及疗效评价密切相关的氨基酸类生物标志物，并通过标志物的时间变化趋势以及代谢途径中标志物之间的相关性挖掘其生物学信息，同时对已确认的生物标志物进行定量分析并建立量化标准。此外，通过生物质控样本与混合标准品溶液的稳定性，确定可控的标准化条件与参数，建立规范化的代谢组学样本质控流程。本研究有望为临床药物肝损伤的诊断与治疗提供重要依据。

联合用药与新药的广泛使用加大了药物性肝损伤发病风险，特异性标志物的缺乏成为药物性肝损伤早期发现和预后判断的瓶颈问题。本研究采用正、负离子检测模式的RRLC-MS/MS 技术结合多变量统计分析方法，针对健康者、药物性肝损伤（肿瘤药物、中药、他汀类降脂药、抗菌药）初诊患者及治疗后转归患者的血清样本开展组学研究，旨在寻找及确认与药物性肝损伤早期诊断及疗效评价密切相关的氨基酸类生物标志物。运用本方法共寻找到114个在肝损伤患者与健康人血清中存在显著性差异的内源性小分子代谢物。同时，结合患者临床的疗效评价，考察可能生物标志物在健康状态、肝损伤初诊病例以及治疗后的三个阶段的动态变化趋势，寻找到了一些与疗效相关的生物标志物。此外，分别建立了上述降脂药、抗肿瘤药及中药引发肝损伤血清样本的多变量数据分析模型，将各致病药物组中影响力较强的可能生物标志物进行比较，获得不同药物引发损伤的共有与特异性生物标志物。本课题的研究结果有望为药物性肝损伤的早期发现、临床疗效评价和预后判断提供重要依据。