Comparative Total Proteome Analysis of NASH for Diagnostic Biomarkers

Abstract

Non-alcoholic fatty liver disease (NAFLD) includes many multifactorial diseases that range from simple to non-alcoholic steatohepatitis (NASH), fibrosis and cirrhosis. It is considered as the one of the major risk factors for chronic liver diseases worldwide. Despite its high occurrence, the underlying molecular mechanisms responsible for the development of NASH remain mostly unknown. In this study, we looked for diagnostic biomarkers for NASH using the omics approach with tissue and serum samples from animals by verifying the obtained results with human serum samples. The tissue samples from control and NASH groups were analyzed by nano liquid chromatography mass spectrometry (nLC-MS). The measured fragments were further analyzed using Proteome Discoverer 2.5 software and identified around 13,000 proteins, with 10,730 common to both groups. Among the identified proteins, 105 highly changed proteins in NASH samples were selected using specific filtering criteria. Multiple Reaction Monitoring (MRM) scanning of selected proteins showed significant expression level changes in tissue and serum samples. Upregulated proteins like annexin2 (ANXA2), galectin 3 (LGALS3) suggested increased membrane trafficking and inflammation, while downregulated metabolic enzymes such as fatty acid synthase (FASN) and glutamate dehydrogenase 1 (GLUD1) indicated potential dysregulation in metabolic pathways in NASH liver tissue. Overall, our findings provide insights into molecular alterations in NASH pathogenesis and highlight potential diagnostic biomarkers.