Objective To investigate the effect of Pulsatilla Saponin D on the metastasis and apoptosis of glioma cells U251MG. Methods The activity of normal astrocytes and glioma cells U251MG was detected by CCK-8 kit. Glioma cells U251MG treated with 0.0, 1.0, 2.0 and 5.0 μmol/L Pulsatilla Saponin D were divided into four groups: PSD group 0.0 μmol/L, PSD group 1.0 μmol/L, PSD group 2.0 μmol/L and PSD group 5.0 μmol/L. Cell migration was detected by scratch test. Transwell detects cell invasion. Hoechst 33258 staining detected apoptosis. MRNA levels were detected by RT-PCR. Protein immunoblot was performed to detect the protein levels of MMP-2, uPA, PAI-1, VEGF, cleaved Caspase-3, cleaved Caspase-9, and c-MET.Results Pulsatilla Saponin D inhibited the activity of normal astrocytes and glioma cells U251MG. Compared with PSD 0.0 μmol/L group, PSD 1.0, 2.0, 5.0 μmol/L group cell migration rate significantly decreased(P<0.01), MMP-2, the uPA, PAI-1, VEGF, c-MET protein levels decreased significantly (P<0.05), the PSD 1 mu number cells invade mol/L group decreased significantly(P<0.05), the PSD 2.0, 5.0 μmol/L group significantly reduced(P<0.01), the apoptosis rate increased significantly(P<0.05), Cleaved Caspase-3 and Caspase-9 protein levels were significantly increased(P<0.05).Conclusions Pulsatilla Saponin D inhibits the migration and invasion of glioma cells U251MG and induces apoptosis by targeting c-MET.