基于UPLC-Q-TOF/MS技术的成人草酸钙尿石症血液代谢组学研究

Blood metabolomics research for calcium oxalate urolithiasis in adults based on UPLC-Q-TOF/MS

目的基于超高效液相色谱-四极杆飞行时间质谱(UPLC-Q-TOF/MS)技术,通过血液代谢组学寻找成人草酸钙尿石症潜在的生物标志物和代谢机制,以指导草酸钙结石的临床诊治。方法前瞻性收集2017年10月—2018年7月北京友谊医院收治的36例双侧上尿路草酸钙结石患者的血液样本并与36例健康体检者的血液样本对照。采用UPLC-Q-TOF/MS技术进行代谢指纹图谱分析。采用主成分分析(PCA)和正交偏最小二乘判别分析(OPLS-DA)对多元数据进行分析。通过对比人类代谢组数据库确定潜在的生物学标记物。最后对所鉴定的化合物行代谢通路分析并构建代谢网络图。结果PCA和OPLS-DA结果显示健康对照者与结石患者血液代谢物存在显著差异。进一步鉴定出包括亚油酸、枸橼酸、尿酸等29种与草酸钙结石患者血液相关的代谢物,以及胆汁酸合成通路、脂肪酸生物合成通路、甘油磷脂代谢通路、鞘脂代谢通路和嘌呤代谢通路等9条血液代谢通路。结论基于UPLC-Q-TOF/MS平台进行血液代谢组学分析,找到了29种成人草酸钙尿石症的潜在血液生物标志物,而嘌呤代谢与脂质代谢可能与草酸钙结石的成因相关。

Objective Based on ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF/MS),the potential biomarkers and metabolic mechanisms of adult calcium oxalate urolithiasis were identified to guide the diagnosis and treatment of calcium oxalate lithiasis.Methods Blood samples of 36 patients diagnosed with bilateral upper urinary tract calcium oxalate stones and 36 healthy controls admitted to our hospital during Oct.2017 and Jul.2018 were collected.Metabolic fingerprints were analyzed with UPLC-Q-TOF/MS.The multivariate data were analyzed with principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA).Potential biological markers were identified by comparing human metabolome datasets.Finally,the metabolic pathways of the identified compounds were analyzed and the metabolic network map was constructed.Results PCA and OPLS-DA showed significant differences in blood metabolites between patients and healthy controls,and identified 29 blood-related metabolites,including linoleic acid,citric acid,uric acid and so on.The most relevant blood metabolic pathways were bile acid synthesis pathway,fatty acid biosynthesis pathway,glycerophospholipid metabolism pathway,sphingolipid metabolism pathway and purine metabolism pathway.Conclusion Blood metabolomics analysis based on the UPLC-Q-TOF/MS platform identified 29 potential blood biomarkers of adult calcium oxalate urolithiasis,and purine and lipid metabolism may be related to the genesis of calcium oxalate stones.