Increased atmospheric nitrogen deposition generally increases plant biomass production until reaching soil N saturation, which could increase the uncertainty of changes in ecosystem temporal stability and their mechanisms. Yet, the response of ecosystem stability to N enrichment and their underlying mechanisms are uncertain, especially when N saturation reached. Here, we conducted a multi-level N addition (0, 2, 5, 10, 15, 25, and 50 g N m-2 year-1; high added rates reached N saturation) experiment from 2018 to 2022 to estimate the effect of simulated N deposition on ecosystem biomass stability in a subalpine grassland located on the Qilian mountain of north-eastern Tibetan Plateau. Our results show that community biomass production increased with an increase in N addition in the first N addition experiment year, but decreased with increase in N addition rates after N saturation in subsequent years. We first find a negative quadratic relationship between biomass temporal stability and added N rate, whereby above N saturation threshold (5 g N m-2 year-1 at this site), increases in N addition reduces biomass temporal stability. The changes in biomass temporal stability are largely determined by dominant species stability, species asynchronous, and species richness. These results provide a better understanding of N-induced effect on ecosystem stability and their underlying mechanisms, which is important to evaluate functioning and services of ecological systems under global change scenarios.