Aligned with the imperatives of national ecological civilization construction, the systematic investigation into the intricate interplay between shifts in land utilization and the assessment of ecosystem services plays a pivotal and indispensable role in advancing ecological civilization. This endeavor holds significant implications. It aids in optimizing the ecological landscape at the regional level and fosters harmonious coexistence between humanity and the natural world. The study utilizes land-use remote sensing interpretation data from three time periods (2000, 2010, and 2020) and employs various methodologies, including equivalent factor coefficient correction, sensitivity analysis, and spatial autocorrelation. The objective is to uncover the spatiotemporal dynamics of land-use changes and Ecosystem Service Value (ESV) in Lanzhou City. Furthermore, geographic detectors are applied to explore the driving factors influencing ESV spatial heterogeneity and their interactions. The research findings indicate the following: (1) From 2000 to 2020, grassland and cropland were the predominant land-use types in Lanzhou City, with cropland and urban land experiencing the most active changes. (2) ESV in Lanzhou City increased from 179.37 billion RMB in 2000 to 193.86 billion RMB in 2020, reflecting an ESV total growth rate of 8.07% and a gradual improvement in the ecological environment. Spatially, ESV exhibits a “west high, east low” distribution pattern, with the center shifting towards the northwest and southeast, gradually reducing spatial imbalance. (3) Analysis of ESV spatial autocorrelation reveals that high-high clusters are predominantly found within the Tulu Gou National Forest Park and the Xinglong Mountain National Natural Reserve, while low-low clusters are primarily concentrated in the central urban area of Lanzhou City. Over the period from 2000 to 2020, the spatial clustering effect of ESV within the study area has progressively intensified. 4)NDVI, precipitation, and GDP emerge as pivotal factors influencing spatial differentiation within Lanzhou City, with natural and societal elements exerting interactive effects on ESV spatial disparities. The research results integrate environmental considerations into the decision-making process, offering valuable insights for formulating targeted ecological protection policies in Lanzhou City. This study embodies concrete measures taken by Lanzhou City in practicing China’s concept of “green water and green mountains are golden silver mountains,” providing a theoretical basis for the harmonious and sustainable development of the ecological economy.