The relationship among multiple ecosystem services and their response to landscape configuration in different scales is a hot issue, which is important to clarify the scale changes of ecosystem services and the regional landscape management. In this paper, Hubei Province was selected as one study area to analyze the responses of four ecosystem services (e.g., water yield, carbon sequestration, crop production and water purification) to landscape configuration at seven spatial scales, including 5 km×5 km, 10 km×10 km, 15 km×15 km, 20 km×20 km, 25 km×25 km, county, and city scales. The multivariate data (e.g., remote sensing data, land use type data, hydrological data, meteorological data, and soil data) were used and analyzed in our study. The results indicated that (1) the spatial distribution of ecosystem services was spatial heterogeneity. The high value area of crop production and low value area of water purification services mainly distributed in central and eastern parts of Hubei Province, respectively. Carbon sequestration and water yield services showed oppositely spatial distributions. (2) The intensity of ecosystem service trade-offs decreased with the spatial scales increasing, while ecosystem service synergy was increased. The change of intensity was more significant than direction among ecosystem services. (3) The impacts of landscape configuration on ecosystem services at the county and city scales were stronger than that at the grid scales. The results demonstrated that landscape configuration had different scale effects on ecosystem services at grid and administrative scales, and the response of ecosystem services to landscape configuration was scale dependence. These findings provide scientific reference for regional ecosystem service protection and landscape pattern regulation.