基于脂类代谢组学研究对乙酰氨基酚对小鼠药物性肝损伤的早期毒性

目的:基于脂类代谢组学研究对乙酰氨基酚(APAP)对小鼠的早期肝损伤毒性,为寻找潜在生物标志物提供参考。方法:将20只小鼠随机分为正常组和APAP肝损伤组,每组10只。APAP肝损伤组小鼠腹腔注射APAP 300 mg/kg建立急性肝损伤模型,正常组小鼠腹腔注射等体积生理盐水。1 h后取血并分离血浆,采用超高效液相色谱串联三重四级杆飞行时间质谱(UPLC-Triple-TOF-MS)法检测小鼠血浆代谢产物进行代谢组学分析,采用主成分分析(PCA)、偏最小二乘法分析(PLS-DA)以及正交偏二乘法分析(OPLS-DA)区分组间代谢轮廓的整体差异,并根据HMDB、Metlin、LIPID MAPS数据库对脂类代谢物进行筛选和鉴定,同时检测小鼠血浆中APAP水平变化。认定OPLS-DA分析中变量权重值(VIP)大于1且P<0.05的脂类代谢物为差异代谢物,并将脂类差异代谢物与血浆中APAP水平进行相关性分析。结果:PCA、PLS-DA以及OPLS-DA结果显示,正常组和APAP肝损伤组样品点位于图形不同的区域,具有良好的区分度。APAP肝损伤组与正常组比较,血浆中5个脂肪酰类代谢产物水平出现显著的升高或降低,8个甘油磷脂类代谢产物水平均显著降低,1个鞘脂类代谢产物水平显著上升;9-硫杂硬脂酸、十四烷二酸、9-过氧化氢-10,12-十八碳二烯酸、三-肉豆蔻酰基肉碱(脂肪酰类)和Scyphostatin A(鞘脂类)水平与血浆中APAP水平显著相关。结论:APAP染毒1 h后血浆脂类代谢组学显示异常,共发现14个相关脂类差异代谢物,其中5个与血浆中APAP水平显著相关。

Study on Early Toxicity of Paracetamol to Drug-induced Liver Injury in Mice Based on Lipid Metabonomics Research

OBJECTIVE:To study early toxicity of paracetamol(APAP)to drug-induced liver injury in mice based on lipid metabonomics research,and to provide reference for finding potential biological marker.METHODS:Totally 20 mice were randomly divided into normal group and APAP liver injury group,with 10 mice in each group.APAP liver injury group was given intraperitoneal injection of APAP 300 mg/kg to establish acute liver injury model;normal group was given constant volume of normal saline intraperitoneally.1 h later,the blood of mice was collected to isolate plasma.UPLC-Triple-TOF-MS method was used to detect plasma metabolites and perform metabonomics analysis.PC A,PLS-DA and OPLS-DA analysis distinguished the difference of metabolism profiles between groups.The lipid metabolites were screened and identified according to HMDB,Metlin and LIPID MAPS databases.Meanwhile,the changes of APAP level in plasma of mice were detected.The lipid metabolites with variable influence in the projection(VIP)greater than 1 and P<0.05 in OPLS-DA analysis were identified as differential metabolites.The correlation between lipid differential metabolites and plasma APAP level was analyzed.RESULTS:PCA,PLS-DA and OPLS-DA results showed that sample points in normal group and APAP liver injury group were located in different areas with good differentiation.Compared with liver injury group and normal group,levels of 5 fatty acid metabolites were significantly increased or decreased;levels of 8 glycerophospholipids were significantly decreased and one sphingolipids was significantly increased.9-thiastearic acid,tetradecanedioic acid,9-hydrogen peroxide-10,12-octadecadienoic acid,L-myristoyl carnitine(fatty acid)and scyphostation A(sphingolipids)levels had a significant correlation with APAP level in plasma.CONCLUSIONS:The plasma lipid metabolomics showed abnormal changes 1 hour after acetaminophen exposure.A total of 14 related lipid differential metabolites are found,and 5 of which are significantly correlated with APAP level in plasma.