Aim With the application of two-dimensional electrophoresis and mass spectrometry to study the effect of tanshinoneⅡA on ox-LDL-induced RAW264.7 mouse macrophage-derived foam cells proteome and explore the molecular mechanism of lipid and anti-atherosclerotic effect of tanshinoneⅡA. Methods The isolated purified human serum LDL, oxidized with CuSO₄ to give ox-LDL, co-incubated with RAW264.7 cells, the formation of foam cells containing tanshinoneⅡA 20 mg/L to continue for 24 hours was used as the tanshinoneⅡA group; and the control group excluding tanshinoneⅡA solution was incubated for 24 hours; cells were disrupted by ultrasonic and 4℃ 12000 g centrifugal 30 min, to get supernatant for protein quantitation; protein of the control group and the tanshinone ⅡA group was loaded with the same amount and two-dimensional electrophoresis (IEF and SDS-PAGE); after the completion of electrophoresis, they were stained with silver nitrate to get proteome map with different samples. Through Labscan difference proteomic analysis software, select the protein with expression difference of more than 2-fold. After gel digestion and mass spectrometry analysis, peptide mass fingerprinting was obtained, by Mascot database searching, combined with the 2-D map of protein molecular weight and isoelectric point information protein identification was realized. Results In TanshinonⅡA treated group calreticulin, vimentin, peroxidase-2, CuZn-SOD, zipper protein, stabilin-1, hematopoietic cell specific protein were up-regulated, while GTP protein, ATP synthesis, mimitin, IL-5, HSP70, translationally controlled tumor protein, chloride ion channel protein were down-regulated. Conclusions TanshinonⅡA improved the lipid regulation of foam cells by decreasing the expression of GTP protein and HSP70 and increasing the expression of CRT. TanshinonⅡA regulated the ability of endocytosis by increasing the expression of stabilin-1 and decreasing the expression of ATP synthesis. It also stimulated the metabolism of lipid and lipoprotein by improving the expression of leucine zipper protein and vimentin. Meanwhile, tanshinonⅡA eliminated ROS and lipid peroxidation by increasing the expressions of peroxidase-2 and CuZn-SOD. It also had the effects of anti-inflammation and anti-apoptosis by reducing the expressions of mimitin and IL-5. TanshinonⅡA played a role of anti-tumor by subsiding the expressions of CLIC1 and TCTP. In conclusion, tanshinonⅡA may have function of lipid regulation and anti-atherosclerosis, and can be used to treat cardiovascular diseases in clinic.