The present study examined the anti-proliferative properties of aqueous (WESM), ethanolic (EESM) and supercritical carbon dioxide (SCESM-5) extracts of Salvia miltiorrhiza in 3T3-L1 adipocytes, and the contents of bioactive compounds salvianolic acid B (SAB), cryptotanshinone (CTS), tanshinone 1 (TS-1) and tanshinone IIA (TS-IIA). Results showed that WESM contained mainly SAB. whereas EESM possessed all the four bioactive compounds, but its concentrations of CTS, TS-1 and TS-IIA were 2-3-fold lower than SCESM-5. SCESM-5 demonstrated the most potent inhibitory effect on 3T3-L1 cell proliferation, and induction of cell apoptosis and cell cycle arrest. Further studies showed that it enhanced intracellular ROS level, down-regulated CD95 (APO-1/CD95), Bcl-2 and PPAR-gamma, up-regulated Bax, AMPK and p-AMPK, and caused activation of caspase-3 and PARP cleavage, as well as JNK and ERK activation and phosphorylation. Pretreatments with antioxidant (Vit E), caspase-3 (z-DEVD-fmk) and ERK (PD98059) inhibitors significantly suppressed the SCESM-5-induced ROS production and cell death. Caspase-3 inhibitor efficiently blocked the Bax expression, caspase-3 activation and PARP cleavage, while ERK inhibitor suppressed the ERK phosphorylation after SCESM-5 treatment. This study concludes that CTS. TS-1 and TS-IIA of SCESM-5 may have contributed to the potent anti-proliferative activity through the MAPK and AMPK pathways in 3T3-L1 adipocytes. (C) 2011 Elsevier Ltd. All rights reserved.