Multi-omics reveals that 4'-O-β-D-glucosyl-5-O-methylvisamminol ameliorates acute liver injury by modulating the miR-30e-3p/Tfrc axis and a multi-dimensional network of ferroptosis, inflammation, metabolism, and gut microbiota.

5-O reduced serum markers of liver injury (ALT and AST) and alleviated liver tissue pathological damage in APAP-induced acute liver injury.

• APAP overuse was used to induce acute liver injury (ALI) in the experimental model.
• 5-O treatment reduced serum ALT/AST levels, indicating hepatoprotective effects.
• Liver tissue pathological damage was alleviated following 5-O intervention.
• Both in vitro and in vivo experiments were conducted to evaluate these effects.

5-O inhibited ferroptosis by downregulating iron uptake transporters and upregulating antioxidant pathway components in cells.

• 5-O downregulated Tfrc (transferrin receptor) and Slc39a14 to reduce iron intake.
• 5-O upregulated Slc7a11 (system Xc- transporter) and GPX4 to enhance antioxidant capacity.
• These effects suggest suppression of the ferroptosis pathway.
• Western blot was among the techniques used to evaluate protein expression.

Whole transcriptome sequencing revealed that 5-O intervention was associated with upregulation of miR-30e-3p and downregulation of Tfrc, suggesting a role in modulating iron metabolism via the miR-30e-3p/Tfrc axis.

• Whole transcriptome sequencing was performed to analyze transcriptome-wide changes.
• 5-O intervention was associated with upregulation of miR-30e-3p.
• Concurrently, Tfrc (transferrin receptor C) was downregulated.
• The analysis also indicated inhibition of the ferroptosis pathway and activation of the PPAR pathway with 5-O treatment.

5-O inhibited the expression of chemokines and inflammatory factors, suppressing inflammatory responses in APAP-induced liver injury.

• 5-O treatment inhibited expression of chemokines and inflammatory factors.
• The anti-inflammatory effect was evaluated using in vitro and in vivo experiments.
• 5-O reshaped intestinal microbiota by inhibiting pro-inflammatory bacterial genera.
• The compound was previously known to have anti-inflammatory and antioxidant activities as a natural coumarin glycoside.

5-O activated the PPARα-Fabp1/Me1 axis to promote lipid metabolism in the context of APAP-induced acute liver injury.

• 5-O activated the PPARα (peroxisome proliferator-activated receptor alpha) pathway.
• Downstream targets Fabp1 (fatty acid binding protein 1) and Me1 (malic enzyme 1) were involved.
• This activation was associated with promotion of lipid metabolism.
• Whole transcriptome analysis confirmed activation of the PPAR pathway with 5-O treatment.

5-O reshaped the intestinal microbiota by inhibiting pro-inflammatory bacterial genera and promoting colonization of the probiotic Lactobacillus.

• 16S rRNA sequencing was used to characterize gut microbiota composition.
• Fecal microbiota transplantation (FMT) was employed as an experimental technique.
• 5-O inhibited pro-inflammatory bacterial genera.
• 5-O promoted the colonization of the probiotic Lactobacillus.

The hepatoprotective effect of 5-O against APAP-induced liver injury involves a multi-dimensional network encompassing ferroptosis, inflammation, metabolism, and gut microbiota.

• The study used a multi-omics approach combining whole transcriptome sequencing and 16S rRNA sequencing.
• Mechanisms were evaluated at multiple levels including cells, tissues, metabolism, and flora.
• The multi-dimensional network was characterized as 'ferroptosis-inflammation-metabolism-flora.'
• The study employed CCK-8, Western blot, whole transcriptome sequencing, 16S rRNA sequencing, and FMT techniques.