Previous research reported litchi thaumatin-like protein (LcTLP) could lead to inflammation, which is a factor causing the adverse reactions after excessive intake of litchi. As a main amino acid in litchi pulp, γ-aminobutyric acid (GABA) was found with anti-inflammatory effect. Therefore, this study aimed to investigate the effects of GABA on LcTLP-induced inflammation through RAW264.7 macrophages and C57BL mice models. In vitro study showed GABA could effectively regulate the level of inflammatory cytokines (interleukin (IL)-1β, IL-6, IL-10, and prostaglandin E2) and Ca2+ in cells, and inhibit the phosphorylation of p65, IκB, p38, c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK). These results indicate GABA alleviated inflammation through nuclear factor-κB and mitogen-activated protein kinase pathway signaling pathways. In vivo experiment was performed to verify the anti-inflammatory effect of GABA, and the results demonstrated that GABA reduced the inflammation and oxidative stress in the liver of LcTLP-treated mice, as it down-regulated the pro-inflammatory cytokines, malondialdehyde, aspartate transferase, and alanine transaminase. The relative expression of phosphorylated p38, JNK and ERK in mice liver with GABA treatment were reduced to 65%, 39% and 80% of the control group, respectively. Furthermore, GABA treatment enriched probiotic bacteria and decreased pathogenic bacteria in mice gut, which reveals GABA could effectively reduce the translocation of gut microbiota.