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The Lactobacillusis of great interest as a natural antioxidant to ameliorate liver injury for its antioxidant capacity. In a previous experiment, Lactobacillus plantarumKLDS 1.0386 was excellent in adhesion and anti-inflammation. This research assessed the antioxidant capacity of L. plantarumKLDS 1.0386 and how it ameliorates the LPS-induced acute liver injury. In vitroresults showed that L. plantarumKLDS 1.0386 had potential antioxidant capacity with 27 genes, and it had better capacities in hydroxyl radicals scavenging and lipid antioxidant than the reference strain L. plantarumWCFS1 (P < 0.05). In vivo results, L. plantarumKLDS 1.0386 significantly reduced hepatic tissue injury, pro-inflammatory cytokines (TNF-α, IL-6), myeloperoxidase (MPO) activities and biochemical indicators of hepatotoxicity (ALT, AST, ALP) (P < 0.05), compared with the LPS-treated mice. Moreover, L. plantarumKLDS 1.0386 increased the relative abundance of the genera, including Lachnospiraceae_NK4A136_group,Muribaculaceae and Alistipes, which significantly increased the level of acetic acid and butyric acid (P < 0.05). The results also showed L. plantarumKLDS 1.0386 significantly adjusted the expression of HO-1, NQO1, CAT, NF-κB, TLR4 and IκB-α in the liver (P < 0.05). Overall, these results indicated that L. plantarumKLDS 1.0386 had great functionality in reducing LPS-induced acute liver injury.