miR ‐204‐5p Promotes Hepatogenic Differentiation of Adipose‐Derived Stem Cells to Ameliorate Acute Liver Failure via the MDM2 / E2F8 / BMP4 Axis

Acute liver failure (ALF) is characterized by extensive hepatocyte necrosis and limited treatment options. Adipose-derived stem cells (ADSCs) show potential for liver regeneration, yet the mechanisms regulating their hepatic differentiation remain unclear. Human ADSCs were infected with lentiviruses to overexpress or silence miR-204-5p, Murine double minute 2 (MDM2), E2F transcription factor 8 (E2F8), or bone morphogenetic protein 4 (BMP4). qPCR, immunofluorescence, and western blot detected mRNA and protein levels of hepatic markers-including hepatocyte nuclear factor 4 alpha (HNF4α), albumin (ALB), and alpha-fetoprotein (AFP). Periodic acid-Schiff (PAS) staining assessed glycogen storage, while enzyme-linked immunosorbent assay (ELISA) measured ALB and urea secretion. Dual-luciferase reporter and co-immunoprecipitation (Co-IP) assays verified molecular interactions and ubiquitination. A Carbon Tetrachloride (CCl₄)-induced ALF model in mice was adopted to investigate the therapeutic potential of miR-204-5p-overexpression ADSCs by fluorescence imaging, histology, tunel staining, and measurement of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels. In vitro, miR-204-5p suppresses MDM2 expression, thereby reducing MDM2-mediated ubiquitination and degradation of E2F8. Stabilized E2F8 transcriptionally activates BMP4, ultimately promoting the hepatogenic differentiation of ADSCs, as evidenced by increased ALB and HNF4α expression, enhanced glycogen synthesis, and elevated urea production. In a CCl₄-induced ALF mouse model, miR-204-5p-modified ADSCs localized to the liver, reduced necrosis and apoptosis, improved hepatic architecture, and decreased ALT and AST levels. miR-204-5p promotes hepatogenic differentiation of ADSCs and ameliorates acute liver failure through the MDM2/E2F8/BMP4 signaling axis, providing a potential molecular target to enhance stem cell-based liver regeneration therapy.