Abstract:　Bacillus　subtilis　has　been　reported　to　maintain　the　homeostasis　of　intestinal　flora.　In　this　study,　a　mouse　model　of　alcoholic　liver　injury　(ALI)　was　constructed　to　study　the　ameliorative　effect　of　B.　subtilis　C10　on　ALI　and　to　further　clarify　its　mechanism　of　action.　Significant　correlations　between　intestinal　flora　and　biochemical　indicators　of　ALI　were　found　by　statistical　correlation　analysis.　Supplementation　with　B.　subtilis　C10　modulated　the　equilibrium　of　gut　flora　by　reducing　the　population　of　detrimental　bacteria　while　enhancing　the　numbers　of　beneficial　microorganisms,　which　resulted　in　an　improvement　in　lipid　metabolism　and　oxidative　stress　in　the　liver.　The　results　of　RT-qPCR　showed　that　B.　subtilis　C10　intervention　regulated　the　main　regulatory　factors　of　liver　lipid　metabolism　(PPAR-α,　SREBP-1c)　and　interfered　with　Nrf-2/Ho-1　signal　pathway,　which　in　turn　ameliorated　alcohol-induced　lipid　metabolism　disorder　and　liver　peroxidation　stress.　In　addition,　liver　metabonomic　analysis　showed　that　B.　subtilis　C10　intervention　reduced　the　production　of　harmful　metabolites　and　increased　beneficial　metabolites　in　the　liver,　thereby　reversing　the　metabolic　disturbances　caused　by　excessive　alcohol　consumption.　KEGG　analysis　showed　that　B.　subtilis　C10　intervention　modulated　liver　metabolic　disorders　and　accelerated　lipid　metabolism　by　regulating　glutathione　metabolic　pathway,　purine　metabolic　pathway,　pantothenic　acid　and　CoA　biosynthesis　pathway,　ABC　transporter　protein　pathway,　and　HIF-1　signaling　pathway.　Taken　together,　these　findings　suggest　that　B.　subtilis　C10　ameliorates　ALI　by　modifying　the　structure　of　intestinal　flora　and　liver　metabolic　pathways　to　attenuate　alcohol-exposure-induced　liver　oxidative　damage　and　lipid　metabolism　abnormalities.　Practical　Application:　Bacillus　subtilis　C10　is　an　effective　probiotic　intervention　that　significantly　ameliorated　alcoholic　liver　injury　in　mice　through　the　gut–liver　axis.　B.　subtilis　C10　can　be　used　as　a　dietary　probiotic　to　develop　functional　foods　with　beneficial　effects　for　the　population　of　excessive　alcohol　consumption.　©　2024　Institute　of　Food　Technologists.