[Objective] The distribution pattern of ecosystem service supply and demand and their main driving factors were analyzed in order to provide a scientific reference for regional ecological protection practices and ecological environmental improvement. [Methods] The supply and demand of five ecosystem services in Hu'nan Province from 2000 to 2020 were assessed, including food production, carbon sequestration, water conservation, soil conservation, and habitat quality by use of the InVEST model, ArcGIS, and other tools to analyze their spatial and temporal evolution patterns and spatial matching relationships, and to determine the driving factors affecting the supply-demand ratios of ecosystem services at the grid scale and county scale using an optimal parameter geodetector model. [Results] ① The total supply of ecosystem services in Hu'nan Province was greater than the total demand. The supply of carbon sequestration services and habitat quality services as well as the demand for water conservation services decreased over time, while the supply and demand of other ecosystem services increased. ② The distribution of supply and demand of various ecosystem services in Hu'nan Province, the clustering characteristics of supply-demand ratios, and the spatial heterogeneity of the spatial matching of supply and demand were obvious. ③ There were some differences in the dominant factors of ecosystem service supply-demand ratios between the two scales, and the influence of socio-economic factors was more significant at the county scale than at the grid scale. The interaction between land use factors and other factors explained the spatial variation of ecosystem service supply-demand ratios more strongly. [Conclusion] The contradiction between supply and demand of ecosystem services at different scales was exceptional, and government agencies at all levels should develop more precise ecological management practices in the future, and use them as a basis to promote the rational use of regional land and to ultimately achieve the goal of regional green development.