Fig. 4

Validation of the inflammatory response and apoptosis as key features in ACLF development. A Heatmap of serum ELISA quantification of immunoinflammatory mediators in rats at different disease stages (ACLF = 16; LC = 3; NC = 3). B Gene profile validation of certain hepatic inflammatory mediators and immune chemokines in liver tissues collected from rats at different disease stages (ACLF = 20; LC = 10; NC = 10). Data are represented as the mean ± SD. * P < 0.05, **** P < 0.0001 vs. NC, Gapdh was used as a reference control gene. C-E Immunoblotting analysis of hepatocellular apoptosis-related markers; β-actin was used as a loading control. F Representative images of the TUNEL assay highlighting the presence of apoptotic hepatocytes (black arrow) in liver tissues collected from rats at different disease stages