基于UPLC-QTOFMS正、负离子模式对无偿献血者乙型肝炎表面抗原阳性血清代谢组学的研究

A Study on the Metabolomics of Serum Hepatitis b Surface Antigen Positive in Unpaid Blood Donors Based on UPLC-QTOFMS Positive and Negative Ion Patterns

目的:基于超高效液相色谱-单四极杆飞行时间质谱(UPLC-QTOFMS)正、负离子模式探讨无偿献血者中乙型肝炎表面抗原阳性和乙型肝炎表面抗原阴性的血清代谢组学的差异,为乙型肝炎的诊断寻找潜在的血清生物标志物。方法:选取2017年10月~2018年1月在青海省血液中心检测的乙型肝炎表面抗原阳性57例(研究组)与同期无偿献血者乙型肝炎表面抗原阴性63例(对照组),利用UPLC-QTOFMS技术建立两组血清代谢指纹图谱,采用主成分分析(PCA)和偏最小二乘法-判别分析(PLS-DA)分析两组间有差异的小分子物质,确定与乙型肝炎相关的生物标志物,并分析相关代谢机制。结果:通过变量重要性投影、质谱鉴定和数据库检索筛选出8个潜在的生物标志物,分别为缬氨酸、胆碱、甘氨鹅去氧胆酸、肉毒碱、高丝氨酸、溶血磷脂酰胆碱、血清溶菌酶和花生四烯酸,涉及胆汁酸代谢、氨基酸代谢、磷脂代谢等。结论:无偿献血者中乙型肝炎表面抗原阳性和乙型肝炎表面抗原阴性的血清代谢物存在显著差异,差异代谢物的发现有助于寻找乙型肝炎的潜在生物标志物,为血液安全提供依据。

Objective: Based on the ultra high performance liquid chromatography-single quadrupole time-of-flight mass spectrometry(UPLC-QTOFMS) positive and negative ions model, the difference of serum metabolomics between positive and negative surface antigen of hepatitis b in unpaid blood donors was discussed, and potential serum biomarkers were found for the diagnosis of hepatitis b. Methods: 57 cases of hepatitis b surface antigen positive(research group) and 63 cases of hepatitis b surface antigen negative in the same period(control group) of unpaid blood donors were selected from October 2017 to January 2018 in qinghai blood center. UPLC-QTOFMS technology was used to establish of serum metabolic fingerprint for two groups. Principal component analysis(PCA) and partial least squares discriminant analysis(PLS-DA) were used to analyze the differences between the two groups of small molecules, determine biomarkers related to hepatitis b, and analyze the related metabolic mechanism. Results: Eight potential biomarkers were filtered by variable importance projection screen, mass spectrum identification and database retrieval, they were valine, choline, glycochenodeoxycholic acid, carnitine, homoserine, lysophosphatidylcholine, serum lysozyme and arachidonic acid, involving the metabolism of bile acid metabolism, amino acid metabolism and phospholipids, etc. Conclusions: There are significant differences in the serum metabolites of positive hepatitis b surface antigen and negative hepatitis b surface antigen in unpaid blood donors, and the discovery of differential metabolites is helpful to find potential biomarkers of hepatitis b and provide basis for the blood safety.