Objective To explore the effect of simulating orthodontic static pressure stimulation on the proliferation of periodontal ligament stem cells and phosphatidylinositol 3-kinase(PI3 K)/Protein kinase B(AKT) signaling pathway. Methods Cell proliferation was measured in vitro culture of periodontal ligament stem cells with good growth conditions under 100 KPa for 1 h, 6 h and 12 h. Apoptosis related proteins and PI3 K/AKT signaling pathway proteins were detected. Results The separated and purified periodontal ligament stem cells had the ability of multidirectional differentiation. The proliferation of the stem cells in the posterior periodontal ligament of 1 h and 6 h was significantly inhibited by pressure treatment, and the expression of apoptotic protein cysteinyl aspartate specific proteinase(Caspase)6 and Caspase9 was significantly enhanced(P<0.01, P<0.05). The expression of PI3 K/AKT pathway protein PI3 K and AKT was significantly enhanced(P<0.01, P<0.05), and was significantly different from the control group(P<0.01, P<0.05), but there was no significant difference between the above indexes after treatment(P>0.05). Conclusion Simulated orthodontic static pressure stimulation significantly inhibits the proliferation of periodontal ligament stem cells, and this process may be related to abnormality of PI3 K/AKT signaling pathway.