Two new compounds (1-2), together with six known ones (3-8), were isolated from the roots of Paeonia lactiflora Pall. The structures of two new compounds were elucidated based on HR-ESI-MS, UV, and NMR spectroscopic data analysis. Moreover, the anti-inflammatory activities of compounds 1-8 were determined using LPS-induced RAW 264.7 cells. Compounds 2, 6, and 8 showed the most potent inhibitory activities on NO production with IC50 values of 17.34 ± 1.5, 27.22 ± 1.3, and 14.79 ± 1.7 μM, respectively. Compounds 4 and 7 showed moderate inhibitory activities. Compounds 3 and 5 showed relatively weak inhibitory activities.