To study the effects of different extracted parts of twotoothed achyranthes root on the proliferation of neural stem cells cultured in vitro. Methods:The systemic solvent method and column chromatography were used to separate the twotoothed achyranthes root into different parts. The effects of various parts of twotoothed achyranthes root on the proliferation of neural stem cells were detected by CCK-8 method, and the optimal parts and concentration was screened. Results:The n-butanol part(25 μg/mL,P<0.01)and the polysaccharide part(12.5 μg/mL, P<0.01)significantly promoted the proliferation of neural stem cells. Conclusion:The n-butanol and polysaccharide part of twotoothed achyranthes root may be the main part for promoting the proliferation of neural stem cells.