基于靶向代谢组学的腹膜透析患者血清氨基酸水平探究

Serum amino acid profiling of peritoneal dialysis patients by target metabolomics approach

目的:分析腹膜透析(PD)终末期肾病(ESKD)患者血清氨基酸谱,探究其氨基酸代谢失衡状况,为氨基酸腹膜透析液的科学合理调整提供参考。方法:回顾性分析2015年1月-2015年6月某院肾内科147例终末期肾病行腹膜透析管植入术患者,与147例同期进行体格检查且未见异常的健康志愿者的完整临床资料。运用超高效液相色谱-串联质谱(UHPLC-MS/MS)法定量检测血清样本中24种氨基酸含量,结合多变量统计学分析筛选腹膜透析患者与健康志愿者的差异代谢物(t检验、变量投影重要性准则)。运用R软件对差异代谢物和实验室检测指标进行相关性分析,探究差异代谢物的生物学功能,并实现PD患者血清氨基酸谱系统聚类。结果:腹膜透析患者与健康志愿者的实验室检测指标与血清氨基酸含量多数存在统计学差异。血清氨基酸定量检测结果的多变量统计分析显示:腹膜透析患者与健康志愿者的氨基酸谱得分图呈现良好的分离趋势。进一步差异代谢物筛选表明,丙氨酸(P<0.001)、马尿酸(P<0.001)等在腹膜透析患者血清中显著升高,而牛磺酸(P<0.001)显著降低,是主要的差异氨基酸。相关性研究表明,不同类型氨基酸(芳香族氨基酸、支链氨基酸等)与实验室检测指标的相关性不同,马尿酸与血肌酐呈现中等强度的相关性(ρ=0.58,P=0.03)。结论:运用UHPLC-MS/MS能够快速、有效识别腹膜透析患者血清氨基酸代谢状况。马尿酸、牛磺酸、丙氨酸3种氨基酸的变化可为氨基酸腹膜透析液的临床合理优化提供参考。

OBJECTIVE To exhibit serum amino acid profiles of peritoneal dialysis patients with imbalanced amino acid metabolism,and provide a reference for rational adjustment of amino acid-enriched dialysate.METHODS Retrospective analysis was conducted on 147 peritoneal dialysis patients as well as 147 health volunteers recruited from a hospital during from January to June,2015.UHPLC-MS/MS method was applied for quantitative analysis of 24 amino acids in human serum.The amino acid profiles of peritoneal dialysis patients and healthy volunteers were analyzed by multivariate statistical analysis,with significant alterations identified through Student-t test and variable importance in projection.A correlation analysis between differential metabolites and laboratory indexes was proposed using R software to explore the biological implications of differentiated metabolites,and system clustering was conducted for an overlook of amino acid profiles in peritoneal dialysis patients.RESULTS Serum amino acid profiles of peritoneal dialysis patients were significantly differentiated from those of healthy volunteers.A distinct separation was observed between the two groups through multivariate analysis.Alanine and hippuric acid were significantly higher（P〈0.001）in peritoneal dialysis patients,while taurine exhibited the opposite trend（P〈0.001）.The three amino acids were annotated as the significant alterations.Different clusters of amino acids（aromatic amino acids,branched chain amino acids,etc.）differed on their correlation with laboratory indexes,and hippuric acid tightly correlated with serum creatinine（ρ=0.58,P=0.03）.CONCLUSION The established UHPLC-MS/MS method is influential in profiling serum amino acids of peritoneal dialysis.Hippuric acid,taurine and alanine should be attended for rational optimization of amino acid-enriched dialysate.