Germinated Foxtail Millet Protein Attenuates CCl4-Induced Acute Liver Injury with Reduced Hepatic Oxidative Stress and Gut Microbiota Remodeling

Foxtail millet (Setaria italica) is a nutrient-rich cereal whose bioactive properties can be enhanced by germination. However, whether germinated foxtail millet protein (GR) pretreatment protects against carbon tetrachloride (CCl4)-induced acute liver injury and how this response relates to the gut microbiota remain unclear. Mice received oral GR at 0.5 or 1 g/kg for 14 days before CCl4 challenge. GR reduced the liver index and serum aspartate aminotransferase and alkaline phosphatase activities, alleviated hepatocellular necrosis and inflammatory-cell infiltration, increased hepatic superoxide dismutase activity, and decreased malondialdehyde content. Transcriptomic analysis of the high-dose GR (HGR) group identified 80 differentially expressed genes relative to the Model group, with enrichment in the cytokine-cytokine receptor interaction, TGF-β signaling, chemokine signaling, and amino acid metabolism. RT-qPCR confirmed higher Cx3cr1 and lower Got1 transcript abundance with HGR. Moreover, 16S rRNA sequencing revealed overall differences in cecal microbial community composition among the Control, Model, and HGR groups. Correlation analysis further linked liver injury and oxidative stress indicators with HGR-associated bacterial genera and hepatic genes. Collectively, GR pretreatment attenuated CCl4-induced acute liver injury and improved hepatic oxidative status. while the HGR-associated transcriptional and microbial changes provide candidate targets for further mechanistic investigation. These findings support further evaluation of GR for potential food applications.