Zeaxanthin, a type of carotenoid, has been proven to exhibit anti-lipogenesis effect; however, the detailed mechanism of this effect is less known. Herein, we evaluated the effects of zeaxanthin on the inhibition of adipogenesis in 3T3-L1 adipocytes and obesity in high-fat diet fed C57BL/6J mice. Zeaxanthin significantly decreased the intracellular lipid content in a dose-dependent manner (5-15 μM) in adipocytes without causing cytotoxicity. In high-fat-diet-induced obese mice, oral administration of 20 mg kg-1 zeaxanthin attenuated the progression of obesity and improved dyslipidemia. It exhibits an anti-adipogenic effect via down-regulating the transcriptional factors and adipocyte-specific genes involved in adipogenesis, both in vitro and in vivo. Furthermore, zeaxanthin treatment reversed the MDI (0.5 mM 3-isobutyl-1-methylxanthine, 1.0 μM dexamethasone, and 1.0 μg mL-1 insulin) and HFD (high-fat diet)-induced inhibition of AMPK phosphorylation in adipocytes and epididymal adipose tissues, respectively, thereby modulating the energy metabolism. These results indicated that zeaxanthin plays anti-adipogenic and anti-obesity roles by inducing AMPK activation, inhibiting lipogenesis, and decreasing intracellular lipid content, adipocyte size, and adipose weight.