基于高效液相色谱-四级杆飞行时间串联质谱的股骨头缺血性坏死患者尿液的代谢轮廓分析

Urinary metabolomics for the osteonecrosis of the femoral head patients based on HPLC-QTOF/MS

摘要:

目的：运用代谢组学的分析研究方法，对股骨头缺血性坏死（osteonecrosis of the femoral head，ONFH）患者尿液进行代谢轮廓分析，从而探索ONFH的分子机制及诊断性生物标志物。方法：运用高效液相色谱-四级杆飞行时间串联质谱技术对26例ONFH患者的尿液（实验组）和26例正常人的尿液（对照组）进行检测，结合主成分分析法（PCA）和正交偏最小二乘法分析（OPLS_DA）对差异代谢物进行模式识别分析并观察组间差异。结果：实验组和对照组的代谢谱得到明显区分，发现并鉴定出28种与ONFH相关的差异代谢物，差异具有统计学意义（P<0.05），其中筛选出二丙基二硫化物、三甲基硒和氨基甲酸作为ONFH潜在的诊断性生物标志物；此外，嘌呤代谢、酪氨酸代谢、果糖和甘露糖代谢的紊乱可能与ONFH发生发展密切相关。生物信息学分析结果显示ONFH患者的骨骼发育、能量代谢、骨代谢、矿物质代谢、脂质代谢、脂质氧化、血管通透性和氧化应激均受到干扰。结论：基于高效液相色谱-四级杆飞行时间串联质谱联合多变量数据统计分析成功运用于ONFH尿液的代谢轮廓分析。

Abstract:

Objective：Osteonecrosis of the femoral head（ONFH），one of the widespread orthopaedic diseases with a decrease in bloodstream to the femoral head，is frequently accompanied by cellular death，trabecula fracture，and collapse of the articular surface. The exactly pathological mechanism of ONFH is unknown. The early and rapid diagnosis biomarkers for this disease remain to explore and further identify. Methods：High performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry（HPLC-Q-TOF/MS） in combination with multivariate statistical analysis was developed and performed to identify the metabolic profiling of urine from 26 ONFH patients and 26 healthy people. Results：Twenty-eight distinctly differential metabolites were detected between the ONFH patients and healthy people. Dipropyl disulfide，trimethylselenonium and carbamic acid were expected to be the potential biomarkers of ONFH. In addition，the up/down-regulation of purine metabolism，tyrosine metabolism，fructose and mannose metabolism were clearly be associated with the ONFH pathogenic progress. The results of bioinformatics analysis suggested a disturbance in energy metabolism，skeletal development，bone metabolism，mineral metabolism，lipid metabolism，lipid oxidation，vascular permeability and oxidative stress of the ONFH patients. Conclusion：Metabolomics could serve as a promising approach for identifying the diagnostic biomarkers and elucidating the pathological mechanism of ONFH.