采用GC-MS代谢组学技术表征阿司咪唑诱导斑马鱼心脏毒性的内源性代谢物

 目的 分析阿司咪唑诱导斑马鱼心脏毒性后内源性代谢物的变化特点,寻找与心脏毒性发生相关的代谢生物标志物。方法 采用阿司咪唑处理发育48hpf(hours post fertilization)斑马鱼幼鱼,观察其心率、静脉窦(sinus venous,SV)-动脉球(bulbus arteriosus,BA)距离、心脏形态等毒性改变,同时采集斑马鱼组织进行气相色谱-质谱联用法代谢组学分析并寻找生物标志物。结果 阿司咪唑处理斑马鱼幼鱼后,心率降低(P＜0.05),出现明显的房室传导阻滞,心脏形态改变,表现出典型的心脏毒性,且呈剂量依赖性。代谢组学分析结果显示,阿司咪唑处理组的代谢轮廓与对照组明显不同,斑马鱼体内葡萄糖、甘氨酸、乳酸肌肝、谷氨酰胺、N-乙酰-L-赖氨酸、L-脯氨酸、柠檬酸、L-酪氨酸、磷酸、胆固醇、棕榈酸等12种代谢物发生了显著变化。结论 这些结果显示,基于新型模式生物斑马鱼的代谢组学技术可以用于表征药物引起的心脏毒性改变,找到的一些与心脏毒性相关的潜在生物标志物将有助于药源性心脏毒性的早期预测和评价。

Endogenous Metabolites Investigation of Cadiotoxicity Induced by Astemizole on Zebrafish Using GC-MS Metabonomics

OBJECTIVE To analysis metabolic changes of zebrafish treated by astemizole, and to find potential cardiotoxicity related biomarkers. METHODS Forty-eight hpf zebrafish was treated by astemizole and cardiac toxicity was denoted with heart rate, sinus venous(SV)-bulbus arteriosus(BA) distance and heart phenotype. Meanwhile, zebrafish tissue samples were obtained and subjected to GC-MS analysis to find potential biomarkers of cardiotoxicity. RESULTS Heart rate of zebrafish treated by astemizole decreased significantly compared with the control groups accompanied with apparent atrioventricular block and cardiac morphological changes. Metabonomics analysis show that the metabolic profiling distinguished astemizole group from the control group and 12 potential biomarkers, glucose, glycine, lactic acid, creatinine, glutamine, N-acetyl-L-lysine, L-proline, citric acid, L-tyrosine, phosphate, cholesterol, palmitic acid, are identified. CONCLUSION These RESULTS showed that metabonomics based on new model organism, zebrafish, can be used to express the astemizole-induced cardiotoxicity. The biomarkers found contribute to the early warning for drug-induced cardiotoxicity.