Abstract
OBJECTIVE: This study was aimed to identify and investigate changes in serum metabolites across the natural progression of chronic hepatitis B (CHB) patients, discover differential metabolites and metabolic pathways in different disease phases, and provide an experimental foundation for clinical diagnosis, staging, treatment, and prognosis of CHB. METHODS: A total of 279 CHB patients hospitalized at Hunan Provincial People's Hospital from June 2020 to March 2021 were enrolled, including 47 in the immune-tolerant (IT) group, 44 in the immune-clearance (IC) group, 73 in the low-replication (LR) group, 57 in the enhanced-replication (ENH) group, 32 in the liver cirrhosis (LC) group, and 26 in the hepatocellular carcinoma (HCC) group, along with 44 healthy controls. Serum metabolites were analyzed using liquid chromatography-mass spectrometry (LC-MS). Preprocessed metabolic data were subjected to univariate and multivariate statistical analyses via MetaboAnalyst 6.0. The metabolites were mapped to the KEGG database to identify key differential metabolic pathways (Impact value > 0.02). RESULTS: The results of principal component analysis (PCA) and Partial least squares discriminant analysis (PLS-DA) pattern recognition showed clustered distribution patterns of metabolites between CHB patients at different natural history phases and healthy controls. Venn diagrams of differential metabolites (DMs) across the six phases of CHB patients revealed that there were phase-specific DMs in each phase: retinyl beta-glucuronide and APGPR Enterostatin in the IT group; creatinine, deoxycorticosterone, O-phosphoethanolamine, and tetradecanedioic acid in the IC group; 4-Pyridoxic acid, sulfasalazine, calcium, and dodecanoylcarnitine in the LR group; oxoglutaric acid, creatine, and 8-Hydroxy-7-methylguanine in the ENH group; umbelliferone, leukotriene D4, malic acid, 3-methylcrotonylglycine, and sulfathiazole in the LC group; 4'-O-methyl-(-)-epicatechin-3'-O-β-glucuronide and DG(16:0/18:1(9Z)/0:0) in the HCC group. All these DMs exhibited AUC values > 0.7, indicating their potential as phase-specific biomarkers for CHB staging. Additionally, compared to healthy controls, there were 6 differential metabolic pathways in the IT group, 8 differential metabolic pathways in the IC group, 6 differential metabolic pathways in the LR group, 7 differential metabolic pathways in the ENH group, 9 differential metabolic pathways in the LC group, and 8 differential metabolic pathways in the HCC group. These differential metabolic pathways were primarily enriched in amino acid metabolism, glycerophospholipid metabolism, hormone metabolism, and energy metabolism. CONCLUSION: Serum DMs and metabolic pathways existed in CHB patients across natural progression phases, demonstrating dynamic variations and potential as staging biomarkers.