Network analysis and experimental validation reveal mitochondrial-mediated programmed cell death mechanisms of Sini...

Acute liver failure (ALF) is a life-threatening syndrome involving multi-organ dysfunction and currently lacks effective pharmacological interventions. Sini Decoction Plus Ginseng Soup (SNRS), a classical Traditional Chinese Medicine herbal formula, has shown protective potential against ALF; however, the mechanistic basis of its action remains incompletely defined.

This study aimed to explore the association between SNRS treatment and mitochondria-related programmed cell death (PCD) regulation in ALF.

A d-galactosamine/lipopolysaccharide (D-GalN/LPS)-induced ALF model was established in Institute of Cancer Research (ICR) mice, and an alpha mouse liver 12 (AML12) hepatocyte injury model was used for in vitro validation. Network pharmacology and molecular docking were employed to identify candidate SNRS-responsive targets and pathways. Mitochondrial function, programmed cell death (PCD)-related markers, and signaling pathway activity were assessed using western blotting, mitochondrial membrane potential and oxygen consumption rate assays, and flow cytometry.

Network analysis suggested phosphoinositide 3-kinase (PI3K)-AKT signaling and necroptosis-related pathways as major candidate pathways potentially responsive to SNRS in ALF. In vivo, SNRS treatment was associated with improved survival, reduced hepatic necrosis, and decreased release of inflammatory damage-associated molecular patterns in d-GalN/LPS-challenged mice, accompanied by modulation of apoptosis- and necroptosis-related markers and preservation of mitochondrial ultrastructure and function. In AML12 hepatocytes, SNRS treatment was associated with enhanced mitochondrial ATP production and attenuation of ATP depletion-related necroptotic features, in parallel with alterations in uncoupling protein 2 (UCP2)-associated oxidative phosphorylation. In addition, SNRS exposure correlated with reduced oxidative stress and apoptotic signaling, concomitant with increased PI3K/AKT1 activation. Notably, these effects appeared to involve distinct but complementary mitochondria-associated regulatory modules rather than a single linear signaling cascade.

Taken together, these in vivo and in vitro findings indicate that SNRS is associated with the modulation of mitochondria-mediated PCD in ALF, potentially involving parallel regulation of PI3K/AKT1-related apoptotic signaling and UCP2/OXPHOS-related necroptotic processes. These observations support the potential of SNRS as an adjunctive therapeutic strategy for ALF, while warranting further mechanistic validation.