In China, Mori Follum, Portulaca oleracea L., and Lycium barbarum L. are traditional Chinese medicine and food used in treating osteoporosis. However, the active compounds, targets, and specific molecular mechanisms of the triangular herb composed of "Mori Follum, Portulaca oleracea L., and Lycium barbarum L." (MPL) have not been fully elucidated. In this study, the chemical components of MPL were first identified by LC-MS. Next, network pharmacology was used to identify the core targets and key pathways of MPL in anti-osteoporosis. And molecular docking simulations were conducted. The prediction results were validated by in vitro experiments using MC3T3-E1 cells. This study screened 22 active compounds from MPL, with a total of 59 targets involved in the treatment of osteoporosis. IL6, TNF, PTGS2, AKT1, ESR1, TP53, HIF1A, PPARG, PRKACA are core targets, and PI3K/AKT is a key pathway. Molecular docking showed that Wongnin, Hirsutine, and Cryptotanshinone strongly bind with AKT1 and ESR1. MPL could promoted MC3T3-E1 cell proliferation and differentiation and regulated the expression of key proteins in the PI3K/AKT/Runx2 signaling pathway. Further, MPL was used as a prebiotic-assisted fermentation to obtain functional yogurt with osteogenic activity. Moreover, MPL1 could improve the fermentation rate, exhibiting better ideal physicochemical properties and sensory quality of yogurt. This study provides a foundation for developing MPL as an anti-osteoporosis functional food.