Frontiers in nutrition
Reduced hepatic GLUL expression accompanies altered RIP3/MLKL-pathway protein abundance in early cholestatic liver injury following bile duct ligation
By Yao B, Xiao F, Tian Z and 1 others.
Abstract
Background
Obstructive cholestasis is characterized by disturbances in bile acid homeostasis, amino acid metabolism, inflammatory responses, and hepatocellular injury. Although RIP3/MLKL-pathway proteins have been implicated in cholestatic liver damage, their relationship with metabolic alterations remains incompletely understood. This study aimed to identify necroptosis-related genes and associated metabolic-pathway alterations in early bile duct ligation (BDL)-induced cholestatic liver injury through integrated bioinformatics analyses and experimental validation.
Methods
Gene expression datasets GSE29776 and GSE166867 were retrieved from the Gene Expression Omnibus database. Differentially expressed genes (DEGs) and necroptosis-related differentially expressed genes (NRDEGs) were identified. Functional enrichment analyses, protein-protein interaction network construction, least absolute shrinkage and selection operator (LASSO) regression, and random forest analysis were performed to prioritize candidate genes. A murine BDL model was established for validation using biochemical assays, histological analyses, RT-qPCR, and western blotting. Phosphorylated RIP3 and MLKL were evaluated both relative to GAPDH and as phosphorylation-to-total-protein ratios.