Ecological well-being, as the material product and utility provided by ecosystem, is closely related to human survival and development, and its scientific calculation is of great significance to regional sustainable development and the improvement of human well-being. In this study, ecosystem contribution rate was used to study regional ecological well-being of Jiangsu province and analyze its spatio-temporal evolution characteristics which was supported by the ecosystem service theory, and then, GeoSOS-FLUS model was used to predict the land use status of Jiangsu province in 2025, while, ecological well-being circulation and spatial autocorrelation were analyzed based on the change of land use from 1995 to 2025. The results show that: (1) From 1995 to 2020, the total ecological well-being and per capita ecological well-being in Jiangsu province decreased with fluctuation, and the eco-economic output efficiency increased by 540.24%. (2) It was predicted that the total ecological well-being of Jiangsu province will increase slightly, with an improvement of 1.05% compared with 2020. Among the 13 study units, only the total ecological well-being of the three cities in northern Jiangsu decreased slightly. (3) During the study period, the profit of ecological well-being in Jiangsu province was generally lower than the loss caused by the change of land use, which can be explained by the results that water ecosystem was an important contribution type to maintain ecological well-being, and the conversion of cultivated land to construction land was the main reason for the spatial loss of ecological well-being in Jiangsu Province. (4) There was a significant spatial negative correlation between eco-economic output efficiency and per capita ecological well-being in Jiangsu province from 1995 to 2025, but the local spatial correlation was not significantly obvious, which was mainly characterized by low-high cluster and high-low cluster, and mainly concentrated in north and south of Jiangsu province. This study can provide theoretical and methodological references for refined management of regional ecosystem and ecological civilization construction.