Auricularia polytricha (AP), an edible mushroom, is continuously being studied due to the medicinal properties. In this study, AP crude extracts from three sequential extraction, starting from hexane (APH), ethanol (APE) and water (APW), were examined for their anti-inflammatory activity and lipid accumulation property in macrophages. APE treatment was found to increase lipid droplet accumulation in both RAW264.7 and LPS-stimulated RAW264.7 cells in a dose dependent manner. Furthermore, nitric oxide production upon LPS stimulation was suppressed on APE pre-treatment. LC-MS analysis was performed to identify the potential bioactive compounds in APE. The PPARγ agonist, 15-Deoxy-Δ12,14-prostaglandin J2-2-glycerol ester (15d-PGJ2-G), was uniquely presented in APE, which was previously described to bind with PPARγ and induces lipid uptake via the upregulation of Cd36. We found that pre-treatment with APE also showed an increase in Cd36 mRNA in RAW264.7 cells, indicating that 15d-PGJ2-G is the potential active compound found in AP. In conclusion, APE exhibited the induction of lipid uptake via CD36, resulting in lipid accumulation.