As a popular medicinal mushroom, Ganoderma lucidum (Leyss. ex Fr.) Karst. (Lingzhi) has been used for centuries in China as food and traditional Chinesemedicine to prevent and treat various diseases. Peroxisome proliferator-activated receptor alpha, gamma and delta (PPARα, PPARγ and PPARδ) are key regulators of energy metabolism. In this study, we presented evidences that G. lucidum spore lipid (GS) dose dependently and preferentially induced the activity of PPARα but not PPARγ and PPARδ. By a gas chromatography–mass spectrometry analysis, we found that GS was rich in unsaturated fatty acids. Additionally, GS induced the expression of PPARa target gene carnitine palmitoyl transferase-1a in human carcinoma HepG2 cells. Collectively, we identified a link between GS and PPARα which suggests that some of the therapeutic effects of GS may bemediated through activating the transcriptional activity of PPARα.