基于高效液相色谱-质谱联用技术的HBV相关性肝纤维化血清代谢标志物筛选

Screening of serum metabolic biomarkers for HBV-related liver fibrosis based on high-performance liquid chromatography-mass spectrometry technology

摘要:

目的： 探讨乙型肝炎病毒（hepatitis B virus，HBV）相关肝纤维化和慢性乙型肝炎（chronic hepatitis B，CHB）患者血清代谢谱差异，寻找与HBV相关肝纤维化的潜在标志物。方法： 采用高效液相色谱-质谱联用技术分析16名HBV相关肝纤维化和16名CHB患者血清代谢谱；应用统计分析及数据库检索，鉴定出2组间差异代谢物；利用MetaboAnalyst 4.0软件对差异代谢物进行代谢通路分析和富集分析，筛选出富集的主要代谢通路及通路上差异代谢物；最后，利用受试者工作特征（receiver operating characteristic，ROC）曲线评价主要差异代谢物的诊断性能。结果： 2组共筛选并鉴定出54种差异代谢产物。代谢通路和富集分析发现，鞘脂代谢通路[-log（P）=9.75，impact=0.42]和甘油磷脂代谢通路[-log（P）=10.90，impact=0.06]是HBV相关肝纤维化的主要富集代谢通路，其中神经鞘磷脂、1-磷酸-鞘氨醇、（神经）鞘氨醇、神经酰胺4种差异代谢物富集于鞘脂代谢通路，而磷脂酰胆碱、磷脂酰丝氨酸、磷脂酰肌醇、磷脂酸、磷脂酰甘油5种差异代谢物富集于甘油磷脂代谢通路；ROC分析显示，9种差异代谢物ROC曲线下面积均大于0.700，均具有良好诊断性能。结论： 鞘磷脂、1-磷酸-鞘氨醇、（神经）鞘氨醇、神经酰胺、磷脂酰胆碱、磷脂酰丝氨酸、磷脂酰肌醇、磷脂酸和磷脂酰甘油诊断HBV相关肝纤维化均具有较高的诊断准确性，可能是HBV相关肝纤维化的潜在代谢标志物。

Abstract:

Objective: To explore the differences in serum metabolic profiles of patients with hepatitis B virus (HBV) -related liver fibrosis and chronic hepatitis B (CHB), and to search for potential biomarkers of HBV-related liver fibrosis. Methods: High-performance liquid chromatography-mass spectrometry was used to analyze the serum metabolic profiles of 16 HBV-related liver fibrosis and 16 CHB patients. By using statistical analysis and database search, the differential metabolites between the two groups were identified. Metabolic pathway analysis and enrichment analysis of differential metabolites using MetaboAnalyst 4.0 software screened out the primary metabolic pathways and differential metabolites on pathways. Finally, the receiver operating characteristic (ROC) curve was used to evaluate the diagnostic performance of major differential metabolites. Results: A total of 54 different metabolites were identified in the two groups. Metabolic pathways and enrichment analysis found that sphingolipid metabolism pathway[-log (P) =9.75, impact=0.42]and glycerophospholipid metabolism pathway[-log (P) =10.90, impact=0.06]were related to HBV-related liver fibrosis mainly enriched in metabolic pathways, including sphingomyelin, 1-phosphate-sphingosine, sphingosine, ceramide and other four different metabolites enriched in sphingolipid metabolism pathways, while phosphatidylcholine, phosphatidyl serine, phosphatidylinositol, phosphatidic acid, phosphatidylglycerol and other five different metabolites were enriched in the glycerophospholipid metabo-lism pathway. ROC analysis showed that the areas under the ROC curve of the nine different metabolites were all greater than 0.700, and all had a good diagnostic performance. Conclusion: Sphingomyelin, 1-phospho-sphingosine, sphingosine, ceramide, phosphatidylcholine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, and phosphatidyl all have a high diagnostic accuracy of HBV-related liver fibrosis, which may be a potential metabolic biomarker for HBV-related liver fibrosis.