Study of ecosystem service supply and demand is an important basis for spatially differentiated control of natural resource development and ecological environmental protection. Research based on the perspective of main function zoning is of great significance for constructing ecological security patterns and promoting sustainable development of regional economy. Taking each county in Hu'nan Province as the research unit, we constructed an Ecosystem service supply and demand coefficient (ESSD). Then, we used the ESSD and the local indicators of spatial association (LISA) to compare two time periods before (2006-2011) and after (2012-2017) the implementation of the main function zoning in Hu'nan Province, aiming to analyze the spatial and temporal variability and spatial correlation characteristics of the supply and demand of ecosystem services. The results showed that:(1) the main function zoning policy had positive guiding effect on protecting the ecosystem and stabilizing the relationship between supply and demand in Hu'nan Province. (2) The ESSD in Hu'nan Province had significantly spatial heterogeneity, showing the spatial distribution characteristics of high in the west and low in the east. There were large differences in the supply and demand of ecosystem services in various functional zones, with key development zones, key ecological functional zones and agricultural production zones mainly showing under-supply, oversupply and balanced supply and demand, respectively. (3) The negatively spatial correlation between supply and demand of ecosystem services in Hu'nan Province had increased, and the difference in spatial equilibrium had intensified.