This study was performed to investigate the lipolytic effects of Portulaca oleracea L. extract in 3T3-L1 adipocytes. The Portulaca oleracea L. was extracted with extrusion method using twin-screw extruder under $58{sim}60rpm$ screw speed, $4{sim}5kg/hr$ feed rate, $140^{circ}C$ extrusion temperature. The lipolytic action of Portulaca oleracea L. extract was estimated by measuring the amount of glycerol and free fatty acids (FFA) released from 3T3-L1 adipocytes and by measuring the cellular lipid content in 3T3-L1 adipocytes. The hormone sensitive lipase (HSL) mRNA level was analyzed using quantitative real-time PCR. The Portulaca oleracea L. extract at 1 to $100{mu}g/ml$ suppressed lipid accumulation. The release of glycerol and FFA into the medium, and the mRNA level of HSL were significantly increased by the addition of Portulaca oleracea L. extract at dose-dependent manner. In conclusion, the Portulaca oleracea L. extract was suggested to have the lipolytic effect through release of lipolytic products (FFA and glycerol) of triacylglyceride to the culture medium and suppression of lipid accumulation via up-regulation of HSL gene expression in 3T3-L1 adipocytes.