Inflammation in adipose tissue is intimately linked with the onset and progression of insulin resistance. The link between insulin resistance and obesity is not completely understood. Resistin, an adipocyte-secreted inflammatory molecule, has been implicated as the link between obesity, inflammation and insulin resistance. During bacterial infection, endotoxin/lipopolysaccharide (LPS) elicits immune and inflammatory responses that can result in a fatal shock syndrome and a series of metabolic alterations and impaired insulin action. LPS has been reported to decrease the expression of resistin mRNA and protein in both lean and db/db obese mice, although the underlying mechanism remains unclear. In the present study, several models such as ex vivo culture of adipose tissues, primary rat adipocytes and 3T3-L1 adipocytes were used to further characterize the effect of LPS on the expression of resistin. We found that acute administration at the doses of 1mg/kg body weight or 80 μg/kg body weight significantly improved the glucose tolerance in mice. Similar results were also observed in mice received a daily intraperitoneal injection of LPS at a dose of 80 μg/kg body weight for 7 days. LPS attenuated both the resistin mRNA and protein in a time- and dose-dependent manner. In the presence of actinomycin D, LPS failed to reduce the half-life of resistin mRNA, suggesting a transcriptional mechanism. The lipid A fraction is crucial for the inhibition of resistin expression induced by LPS. Polymyxin B (PMB), which prevents the activation of toll-like receptor 4 (TLR4) by LPS, significantly attenuated the inhibitory effect of LPS on resistin expression in primary adipocytes, 3T3-L1 adipocytes and ex vivo adipose tissues, indicating that LPS activates TLR4 to inhibit the expression of resistin in adipocytes. Pharmacological intervention of c-Jun N-terminal kinase (JNK) reversed the inhibitory effect of LPS. LPS down-regulated CCAAT/enhancer binding protein α (C/EBP-α) and peroxisome proliferators activated receptor γ (PPAR-γ), while activation of C/EBP-α or PPAR-γ by either over-expressing these transcriptional factors or by rosiglitazone, an agonist of PPAR-γ, blocked the inhibitory effect of LPS on resistin. C/EBP homologous protein 10 (CHOP 10) was up-regulated by LPS, while a CHOP 10 antisense oligonucleotide reversed the decrement of resistin protein induced by LPS. Taken together, the present study demonstrates that inflammation induced by LPS can decrease the resistin expression in adipose tissues. LPS inhibits resistin expression through a unique signaling pathway involving TLR4, JNK, CHOP 10, and C/EBP-α/PPAR-γ.