Assessing the impact of urbanization on ecosystem services is an important part of improving understanding of the harmonious coexistence between people and nature. Despite recent progress, it remains challenging for influence mechanism of different urbanization stages and areas on ecosystem services and their trade-offs and synergies. Here, we analyzed the responses of four ecosystem services (crop production, water retention, heat mitigation, flood mitigation) and their trade-offs and synergies to the spatial and temporal differentiation of urbanization in the Su-Xi-Chang region of China from 1990 to 2020. The results showed that the relationship between ecosystem services and urbanization was "U"-shaped curve. In terms of space and time, the greatest loss of ecosystem services occurred in developing urban areas and during the rapidly urbanizing stage. Also, urbanization weakened trade-offs and enhanced lose-lose situations, showing an "S"-shaped curve. Based on the results, some suggestions are proposed aimed at promoting sustainable urban development.