Screening Of HBV-related Liver Fibrosis Serum Metabolic Markers Based On High-performance Liquid Chromatography-tandem Mass Spectrometry

Objective: To investigate the differences in serum metabolomes between patients with hepatitis B virus(HBV)-associated liver fibrosis and chronic hepatitis B(CHB),and to find potential markers associated with HBV-associated liver fibrosis.Methods: Serum metabolism profiles of 16 HBV-associated liver fibrosis and 16 CHB patients were analyzed by high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS).Principal component analysis(PCA),variable projection importance(VIP),other statistical analysis,and database search were applied to screen and identify differential metabolites.Pathway and enrichment analysis were performed using Metabo Analyst software(version?)to screen for differential metabolites enriched in major metabolic pathways,and the receiver operating characteristic(ROC)curve was used to evaluate the diagnostic performance of key differential metabolites.Results:(1)A total of 54 endogenous differential metabolites,such as sphingomyelin,1-phospho-sphingosine,phosphatidylcholine,phosphatidic acid,phosphatidylglycerol,phosphatidylserine,phosphatidylinositol,uric acid,Threonine,lactic acid,deoxycholic acid,Cinnamic acid,Carnitine,Sphingosine,ceramide,tyrosine,and Mannitol,were selected and identified.(2)Metabolic pathway and enrichment analysis revealed that four different metabolites,sphingomyelin,1-phospho-sphingosine,(neuro)sphingosine,and ceramide,are mainly enriched in the sphingolipid metabolic pathway(-log P=9.747,impact=0.420).Moreover,five differential metabolites,such as phosphatidylcholine,phosphatidylserine,phosphatidylinositol,phosphatidic acid,and phosphatidylglycerol,are mainly enriched in the glycerophospholipid metabolic pathway(-log P=10.899,impact=0.063).Ethanolphosphate is present in both the lipid metabolism pathway and the glycerin phospholipid metabolism pathway.(3)ROC curve analysis showed that,phosphatidylglycerol(PG)has an area under the curve(AUC)of 0.914,a sensitivity of 87.5%,a diagnostic specificity of 93.75%,and a positive predictive value of 93.33%,and a negative predictive value of 88.24%;Sphingosine-1-phosphate has an AUC of 0.781,a sensitivity of 75%,a diagnostic specificity of 81.25%,and a positive predictive value of 80%,and a negative predictive value of 76.47%;Phosphatidylserine has an AUC of 0.883,a sensitivity of 81.25%,a diagnostic specificity of 87.5%,and a positive predictive value of 86.67%,and a negative predictive value of 82.35%;Phosphatidic acid has an AUC of 0.756,a sensitivity of 68.75%,a diagnostic specificity of 75%,and a positive predictive value of 73.33%,and a negative predictive value of 70.59%;Ceramide has an AUC of 0.82,a sensitivity of 93.75%,a diagnostic specificity of 75%,and a positive predictive value of 78.95%,and a negative predictive value of 92.3%;Sphingomyelin has an AUC of 0.855,a sensitivity of 87.5%,a diagnostic specificity of 81.25%,and a positive predictive value of 82.35%,and a negative predictive value of 86.67%;Phosphatidylcholine has an AUC of 0.813,a sensitivity of 81.25%,a diagnostic specificity of 62.5%,and a positive predictive value of 68.42%,and a negative predictive value of 76.92%;Phosphatidylinositol has an AUC of 0.879,a sensitivity of 87.5%,a diagnostic specificity of 87.5%,and a positive predictive value of 87.5%,and a negative predictive value of 87.5%;(Nerve)Sphingosine has an AUC of 0.834,a sensitivity of 81.25%,a diagnostic specificity of 81.25%,and a positive predictive value of 81.25%,and a negative predictive value of 81.25%.Conclusion: There were Significantly different metabolites in metabolic profiles between the groups of patients with hepatitis B liver fibrosis and chronic hepatitis B patients.Nine major differential metabolites enriched in the sphingolipid metabolism pathway and the glycerophospholipid metabolism pathway have good performance in the diagnosis of hepatitis B fibrosis,They are potential metabolic indicators of hepatitis B fibrosis,and May play an important role in the occurrence and development of hepatitis B liver fibrosis.