AIM: To investigate the effect of Astragaloside Ⅳ (As-Ⅳ) on low-glucose mediated tumor immunosuppression microenvironment and its molecular mechanism. METHODS: MTT assay was used to detect the effect of As-Ⅳ on the proliferation of CD4+T cells in low-glucose microenvironment in vitro. By ELISA experiment and qPCR detection of interleukin 2 (IL-2), interferon - gamma (IFN-γ), CD40L and transforming growth factor beta 1 (TGF-β1) level; Western blot was used to detect the expression of glucose transporter 1 (Glut-1), key glycolytic enzymes (HK, PFK1 and PK), AKT/mTOR signaling pathway and AKT/GSK3β signaling pathway in CD4+T cells. Molecular docking and AKT inhibitor experiments were used to verify the results. B16-PKM2-OE was used to establish a low-glucose tumor microenvironment animal model for verification. RESULTS: MTT assay showed that As-Ⅳ promoted the proliferation of CD4+T cells in low-glucose microenvironment (P<0.05). The results of ELISA and qPCR experiments showed that As-Ⅳ could increase the levels of IL-2, IFN-γ and CD40L, and reduce the level of TGF-β1 in tumor tissues (P<0.05). Western blot results showed that As-Ⅳ promoted Glut-1 protein expression on the surface of CD4+T cells, up-regulated the expression of glycolysis key enzymes, and activated AKT/mTOR and AKT/GSK-3β signaling in a concentration-dependent manner. Molecular docking and join AKT inhibitors As the experiment results indicate-Ⅳ activated AKT/mTOR signaling and AKT/GSK-3β signal; Animal experiments showed that As-Ⅳ exerted anti-tumor effect by activating the proliferation and activation of CD4+T cells in low-glucose microenvironment. CONCLUSION: As-Ⅳ promote sugar by activation of AKT/Glut signal micro environment of CD4+T cell proliferation and activation play a role of anti-tumor.