Supplementary file 1_Hepatoprotective Formula 919 ameliorates hepatic steatosis and metabolic disturbance in NAFLD with involvement of AMPK signaling.docx

HF919 treatment increased hepatic p-AMPK abundance and Ppara expression and reduced the expression of Tlr4, Nfkb2, and Tgfb1. Hepatic F4/80-positive staining was significantly reduced, accompanied by increased expression of the macrophage-associated genes Mertk and Arg1. Public liver transcriptomic analyses showed lower AMPK- and fatty-acid-oxidation-associated transcriptional features in NASH and in more severe histopathological categories, supporting the disease relevance of these metabolic alterations.

Integration of HF919-predicted targets with disease-related transcriptomic changes identified metabolic and injury-related candidate pathways, while molecular docking showed favorable predicted docking scores for selected ligand–target pairs. Exploratory SCFA analysis showed shifts in several fecal SCFAs toward the control-associated pattern and associations with selected hepatic metabolic and molecular parameters.</p>Conclusion<p>HF919 ameliorated hepatic steatosis and metabolic abnormalities in HFD-induced NAFLD rats.

The accompanying increase in hepatic p-AMPK abundance and Ppara expression, together with disease-context evidence from human liver transcriptomic datasets, supports the possible involvement of AMPK-associated metabolic regulation. Changes in selected injury-related and macrophage-associated markers and fecal SCFA profiles provided additional secondary or exploratory observations. Further mechanistic studies are needed to establish the causal contribution of AMPK signaling to the effects of HF919.</p>