The negative impacts of rapid urbanization have become key factors in threatening regional ecological security and undermining human well-being. The rapid expansion of construction land and high population concentration accompanying urbanization can drastically alter the structure and function of ecosystems in a short timeframe, thereby affecting the supply of ecosystem services and severely damaging ecosystem stability. However, the impact of urbanization on ecosystem services remains uncertain and is characterized by non-linear relationships, yielding effects that may range from highly significant to negligible. Therefore, a clear understanding of how urbanization affects ecosystem services (ESs) can provide more comprehensive information to support ecological governance decisions. In this study, the spatial and temporal distributions of four ESs, namely water yield (WY), soil conservation (SC), nitrogen export (NE), and habitat quality (HQ), and four factors of urbanization, namely construction land percentage, economic density, population density, and nighttime lighting, were analyzed in the Dongting Lake Basin from 1990 to 2019. Moreover, this study explored the spatial effects of urbanization on ecosystem services using bivariate spatial autocorrelation, geographically weighted regression, and spatial econometric models. The results showed that: (1) Water yield, soil conservation, and nitrogen export increased by 19.54%, 3.96%, and 0.78%, respectively, while habitat quality decreased by 0.016. The urbanization indicators showed continuous growth, and the spatial patterns of changes in various urbanization indicators were highly similar. (2) Significant spatial correlations were observed between urbanization and ecosystem services, with distinct local clustering characteristics. (3) The impact of urbanization on ecosystem services exhibited pronounced spatial heterogeneity and nonlinear driving characteristics. The direction and magnitude of these impacts changed over time, with their overall effects gradually diminishing. Urbanization showed a positive impact on WY and a significant negative impact on HQ. For SC and NE, the construction land percentage (CLP) showed a significant positive effect, while the effects of the nighttime light data (NLD) shifted from predominantly negative to positive. Meanwhile, the negative effects of economic and population densities expanded significantly in terms of their spatial extent. (4) Urbanization effects displayed clear spatial spillover, impacting and receiving feedback from surrounding areas. Moreover, the driving efficacy of different urbanization indicators showed considerable individual variation and temporal dynamics. The intensity of both the direct and indirect effects of urbanization showed a weakening trend. This study provided comprehensive insights into the spatial effects of urbanization on ESs, and these findings offer a reference for urban planning decisions and for promoting habitat improvements that balance economic development with ecological protection.