黄芪口服液降低大鼠顺铂毒性作用机制的尿液代谢组学研究

Untargeted Urinary Metabolomic Study on Toxicity-alleviation Effect of Huangqi Oral Solution in Cisplatin-exposed Rats

采用基于液相色谱-飞行时间质谱联用(LC-TOF-MS)技术的代谢组学方法,分析大鼠尿液内源性代谢物的变化,研究黄芪口服液(HO)降低大鼠顺铂(CDDP)毒性的作用机制。采用低剂量多次腹腔注射CDDP的方法建立CDDP染毒大鼠模型,并连续给予16天HO。于第18天收集正常对照(Control)组、顺铂模型(CDDP)组和黄芪口服液(HO)组大鼠的24 h尿液,进行LC-TOF-MS分析,以获取尿液代谢物组数据集,对所得数据进行主成分分析(PCA)和正交偏最小二乘法-判别分析(OPLS-DA)等多元统计分析,以筛选潜在生物标志物。于第20天采集大鼠血清测定肌酐和尿素氮水平。血清指标测定结果表明,HO可以显著降低CDDP染毒大鼠的肌酐和尿素氮水平(p<0.05)。PCA得分图显示,3组可分别聚类,HO组位于Control组和CDDP组中间,表明HO可部分改善CDDP所致大鼠尿液代谢产物的异常变化。综合OPLS-DA分析、t检验和倍数变化分析结果,最终共筛选并初步鉴定出35个尿液代谢产物作为HO减毒相关的潜在生物标记物。代谢通路分析结果表明,HO可通过纠正体内氨基酸代谢、能量代谢和核苷酸代谢等通路的紊乱,降低CDDP所致机体毒性。

A liquid chromatography-quadrupole-time-of-flight mass spectrometer(LC-Q/TOF-MS)basedurinary metabolomic approach was employed to assess the toxicity-aneviation effect of Huangqi oral solution(HOs)on cisplatin-exposed rats and explore its possible mechanisms.Rat toxicity model was developed bymultiple intraperitoneal injection of low-dose cisplatin,while HOs was orally administrated to ratssimultaneously for16consecutive days to attenuate or reduce the cisplatin-induced toxicity.24-hour urinesamples on day18were collected and analyzed using LC-Q/TOF-MS to obtain the dataset of urinarymetabolites.Principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-D A)were employed to assess the quality of the dataset and screen the potential toxicity-alleviationbiomarkers.The serum level of rat creatinine and urea nitrogen on day20was determined,and the resultsshowed that successive administration of HOs significantly reduced the cisplatin-induced increase of creatinineand urea nitrogen.PCA cluster analysis clearly demonstrated that HOs could partly improve the CDDP-inducedabnormality of metabolic profiling.35urinary metabolites were finally screened as the potential biomarkersassociated with the toxicity-attenuation effect of HOs,according to the combination of the analysis results ofOPLS-D A,t-test and fold change analysis.Further metabolic pathway analysis revealed that HOs could restorethe metabolic disorders of amino acid,energy and nucleotide,thereby exerted its toxicity-alleviation effect.