Abstract: The purpose of the present study was to investigate the anti-proliferative and apoptotic effects of tetramethypyrazine(TMP)on HepG2 and to elucidate the underlying mechanisms. CCK-8 was introduced to analyze the HepG2 cells proliferation. Cell apoptosis, mitochondrial membrane potential(ΔΨ_m)and cytochrome C were measured by high content screening(HCS). Cleaved-caspases protein expression was detected by Western blot. CCK-8 assay indicated that TMP significantly inhibited HepG2 cells proliferation in dose-dependent manner compared with the control group. Moreover, it was found that TMP could also induce HepG2 cell apoptosis, directly increase the release of cytochrome C, decrease ΔΨ_m and increase cleaved-caspase-3 and cleaved-caspase-9 protein expression. TMP may inhibit cell proliferation and induce cell apoptosis by stimulating the mitochondrial pathway apoptosis in HepG2 cells.