基于UPLC/QTOF-MS的代谢组学研究辛伐他汀对不同性别小鼠尿液中代谢物的影响

UPLC/Q-TOF MS-based metabolomics to study the effects of simvastatin on urine metabolites in mice of different genders

目的:基于代谢组学技术研究辛伐他汀对小鼠尿液中脂类代谢物的影响。方法:收集给予药物辛伐他汀后不同天数的小鼠尿液样品,采用超高效液相色谱-高分辨飞行时间质谱(UPLC/Q-TOF MS)联用,Acquity BEH C_(18)色谱柱,流动相为水-乙腈,梯度洗脱,电喷雾离子源,负离子检测模式,通过数据采集进行代谢轮廓分析。采用MarkerLynxXS软件读取总离子流色谱图中峰信息,利用主成分分析和正交偏最小二乘法-判别分析对样品进行分类。筛选出候选的差异代谢物后,通过一级、二级质谱碎片信息,比对数据库鉴别差异代谢物的结构。结果:给药辛伐他汀后,雌鼠和雄鼠的体质量增长速率存在差异,尿液中的代谢物变化有所不同。鉴定出棕榈酸、胆甾烷五醇、羟睾酮葡糖醛酸苷、硝基亚油酸胆固醇、磷脂酸和磷脂酰乙醇胺共6个脂类代谢差异物,呈现出对体内脂类代谢的影响,在雌雄个体中呈现不同的变化规律。结论:研究结果提示辛伐他汀的临床用药需要考虑性别差异的因素。

Objective:To study the effects of simvastatin on the lipid metabolism of urine samples in mice by metabolomics.Methods:Urine samples from mice exposed to different doses of simvastatin were collected.The samples were analyzed by ultra-performance liquid chromatography/high resolution time-of-flight mass spectrometry(UPLC/Q-TOF MS)using Acquity BEH C18 column,water-acetonitrile gradient eluted mobile phase,electrospray ion source and negative ion mode.The Markerlynxs software was used for collecting peak information of total ion chromatograms.The acquired data was processed by principal component analysis and orthogonal partial least squares-discriminant analysis.After screening for candidate differential metabolites,the structure of the differential metabolites was identified by the information of MS and MS/MS spectra.Results:After the administration of simvastatin,there was a difference in the rate of weight gain between female and male mice.And their metabolites also showed variation tendency.Six differential lipid species were identified,including palmitic acid,cholestane-pentol,oxo-androstenrone-glucuronide,cholesteryl nitrolinoleate,phosphatidic acid,and phosphatidylethanolamine,showing the changes of lipid metabolism and different patterns of variation in male and female individuals.Conclusion:The results suggest that the clinical use of simvastatin requires the consideration of gender differences.