Abstract: Objective: To explore the molecular mechanism of water extract and alcohol extract of ginseng and mulberry in treating osteoporosis by means of network pharmacology. Methods: The effects of water extract and alcohol extract of ginseng and mulberry on proliferation rate and testosterone secretion of mouse leydig cells (TM3), and the effects of water extract and alcohol extract of medicine pair on proliferation rate and alkaline phosphatase (ALP) activity of osteoblasts (MC3T3-E1) were observed. Collect chemical components, components and disease-related targets of ginseng and mulberry respectively through traditional Chinese medicine composition database TCMSP, GeneCards and OMIM. After the intersection of that, using STRING database to analyze protein interaction among key targets, and using DAVID database to analyze biological functions and pathways. The related results were mapped and analyzed by Cytoscape 3.7.0. Autodock software was used to perform molecular docking between potential pharmacodynamic substances and key targets. Results: Compared with the model control group, the cell proliferation rate of TM3 administration group was significantly higher than that of single drug, and it was concentration-dependent. The highest proliferation rate was found at the concentration of 200 μg/mL, which was 97.31% and 91.67% (P<0.01), and testosterone secretion was 3.48 and 3.06 ng/mL. Compared with the blank control group, the cell proliferation rate of MC3T3-E1 administration group increased significantly. The proliferation rate of cells at the concentration of 200 μg/mL was the highest in the group of water extract administration, and the proliferation rate was 155%. The highest cell proliferation rate was 142.8% at 100 μg/mL of alcohol extract. The results of ALP analysis showed that the drug had obvious increasing effect on both water extract group and alcohol extract group. In this study, 28 active components and 69 key targets were collected, and 257 biological process, 34 cell compound and 62 molecular function were collected by GO function enrichment. 109 pathways were collected from KEGG pathway enrichment. The results of molecular docking showed that the energy of potential pharmacodynamic substances docking with key targets IL6, ALB, MAPK8 and CASP3 was lower than 0 kcal/mol. Conclusion: This paper revealed the potential pharmacodynamic substances and targets of ginseng-mulberry medicine in the treatment of osteoporosis, which would lay a foundation for the development and follow-up research of ginseng-mulberry medicine.