In this paper, 3-acetylaconitine and songorine were extracted and separated from the total alkaloids of Aconitum szechenyianum Gay., and their physiological activity on inhibition of human liver cancer cells were studied by MTT method to study the cytotoxic patterns of drugs, cell morphology experiment to observe cell morphology, flow cytometry to analysis cell apoptosis and cell cycle, and laser confocal microscopy to analysis cell apoptosis. The antitumor experiments of 3-acetylaconitine and songorine showed that the antitumor effects were gradually enhanced with the increase of the concentration. The number of cells was significantly reduced, and the growth of HepG2 was inhibited significantly after the drugs act on HepG2. In addition, the ratio of cells early apoptosis and late apoptosis increased, and the G1 phase cells gradually increased, but G2 and S phase cells gradually decreased with the prolonging of treatment time. The antitumor research of 3-acetylaconitine and songorine in vitro had confirmed their effects on inhibition tumor cell proliferation and induction liver cancer apoptosis, which showed that 3-acetylaconitine and songorine could become the new cell apoptosis inducers of liver cancer, and laied the foundation for new antitumor drug lead compounds.