OBJECTIVE To screen the optimal compatibility ratio of Gentiana straminea Maxim.(G.S Maxim) and Rhodiola crenulata (Hook. f. et Thoms.) H. Ohba(R. crenulata), and explore its anti-hypoxia effect and possible mechanism through in vivo and in vitro experiments and network pharmacology. METHODS PC12 cells were divided into groups, and the cell hypoxia model was established by Na2S2O4, H2O2 and physical hypoxia methods, and the compatibility ratio of G.S Maxim-R. crenulata was initially screened. Eighty SPF male Kunming mice were randomly divided into blank group, model group, positive group, and different compatibility ratio groups of G.S Maxim-R. crenulata(2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2), with 8 mice in each group. Gastric drug delivery in 15 d in advance, in addition to the blank group, the rest of the groups were normal pressure hypoxia experiments. Groups of mice survival time was recorded. Detected the levels of inflammatory cytokines, MDA content, and SOD activity in the lung tissues of mice in the blank group, model group, positive group, G.S Maxim-R. crenulata 4:6 group and 3:7 group. The anti-hypoxia mechanism of G.S Maxim-R. crenulata was investigated by network pharmacology and molecular docking, and verified by qPCR. RESULTS In vitro experiments showed that G.S Maxim-R. crenulata had better anti-hypoxia activity. In vivo experimental results showed that the combination ratio of G.S Maxim-R. crenulata with 4:6 and 3:7 could significantly improve the survival time of mice, reduce the contents of NF-κB, IL-6, IL-1β and MDA in lung tissue, and increase the content of IL-10 and SOD activity, and the effect of G.S Maxim-R. crenulata 3:7 group was the best. Network pharmacological studies showed that the potential active components of G.S Maxim-R. crenulata in anti-hypoxia might be ursolic acid, oleanolic acid, and ethyl gallate, etc. The core targets included SRC, PIK3CA, MAPK3, etc., and its anti-hypoxia signaling pathways mainly included PI3K-Akt, HIF-1, etc. The results of qPCR showed that G.S Maxim-R. crenulata could increase the expression of PI3K, Akt, mTOR and p62 in the lung tissue of hypoxic mice. Molecular docking verification showed that the core targets SRC, PIK3CA, and p62 had good binding activity with potential active components such as oleanolic acid, kaempferol, ethyl gallate and quercetin. CONCLUSION G.S Maxim-R. crenulata has anti-hypoxia activity, which may be related to PI3K/Akt signaling pathway through anti-inflammatory, anti-oxidative stress and regulation of autophagy.