Identification of overexpressed cytokines as serum biomarkers of hepatitis C virus-induced liver fibrosis using bead-based flexible multiple analyte profiling

Hepatic inflammation is the stimulator to activate hepatic stellate cells (HSCs) and triggers fibrogenesis. Cytokines are produced during liver inflammation and maybe considered as liver fibrosis biomarker. The aim of this study was to investigate whether cytokines can be used as reliable biomarkers of liver fibrosis using flexible multi-analyte profiling (xMAP). A total of 61 chronic hepatitis C patients with different severity of liver fibrosis were enrolled. Liver biopsy was used as standard to assess the severity of fibrosis according to METAVIR classification. Afterward, 15 samples from healthy controls were analyzed and totally 50 cytokines were screened using flexible multi-analyte profiling to discover differential biomarkers. Finally, levels of protein expressions of individual stages of liver fibrosis were measured. In histological examination, the necroinflammatory score (histology activity
index, HAI) was increased from F1 to F4 stage in hepatitis C virus (HCV) infected patients, indicating that inflammation was accompanied with the progression of liver fibrosis. Using flexible multi-analyte profiling, four serum cytokines, including IFN-α2 (p=0.023), GRO-α (p=0.013), SCF (p=0.047) and SDF-1α (p=0.024), were identified under antibody specific recognition and elevated with HAI score. This study reveals the relationship between cytokines and liver fibrosis, and demonstrated that IFN-α2, GRO-α, SCF and SDF-1α may be used as biomarkers to predict liver fibrosis. The overexpressed cytokines may play a role in the progression of liver fibrosis and deserves further investigation.