Soil moisture (SM) significantly affects the exchange of land surface energy and the stability of terrestrial ecosystems. Although some conclusions have been drawn about the effects of SM on the ecosystem water use efficiency (WUE), the influence mechanism and the quantitative assessment framework of SM on WUE are still unclear. This study provides an analysis framework for the feedback relationship between SM and WUE based on the dependence of the evaporation fraction on SM and output datasets from remote sensing and the Global Land Data Assimilation System. The results show that the range of WUE of terrestrial ecosystems of China was 0.02–19.26 g C/kg H2O in the growing season with an average value of 1.05 g C/kg H2O. They also show a downward trend in 43.99% of the total area. In the evapotranspiration (ET) pathway, SM negatively affected WUE, and the sensitivity coefficient ranged from −18.49 to −0.04. In the net primary production (NPP) pathway, the sensitivity coefficient ranged from −68.66 to 43.19. Under the dual effects of the ET and NPP pathways, the influence of SM on WUE was negative in 84.62% of the area. Variation in SM led to significant WUE variability. Generally, the percentage change in WUE (ΔWUE) ranged from 0% to 190.86%, with an average value of 28.02%. The maximum ΔWUE ranged from 0% to 758.78%, with an average value of 109.29%. The WUE of forest ecosystems showed strong resistance to SM variation, whereas that of non-forest vegetation was more sensitive to SM variation. This analytical framework provides a new perspective on the feedback relationship between WUE and SM in terrestrial ecosystems.