Cudrania tricuspidata fruit has anti-inflammatory and anti-obesity effects. The present study aimed to determine the effect of C. tricuspidata fruits vinegar (CFV) on lipopolysaccharide (LPS)-induced adipocytes inflammation in 3T3-L1 adipocytes. The differentiation of 3T3-L1 adipocytes was induced by dexamethasone (DEX), 3-isobutyl- 1-methylxanthine (IBMX) and insulin. Differentiated 3T3-L1 adipocytes were incubated with CFV followed by LPS treatment. The secretion of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1) were measured using ELISA. The expression of toll-like receptor 4 (TLR4), myeloid differentiation primary response 88 (MyD88), tumor necrosis factor receptor-associated factor 6 (TRAF6) and nuclear factor kappa B (NF-κB) were measured by western blotting. The results showed that CFV dose-dependently inhibited lipid droplet generation and LPS-induced TNF-α, IL-6 and MCP-1 production in the adipocytes. CFV inhibited NF-κB activation, and the LPS-induced expression of TLR4, MyD88, and TRAF6. These results indicated that CFV inhibits the LPS-induced inflammatory response in adipocytes by inhibiting TLR4 signaling, including NF-κB pathways.