Separating the impacts of climate change and human activity on actual evapotranspiration (ET) is important for reducing comprehensive risk and improving the adaptability of water resource systems. In this study, the spatiotemporal distribution of actual ET in the Aksu River Basin, Northwest China, during the period 2000–2015 was evaluated using the Vegetation Interfaces Processes model and Moderate Resolution Imaging Spectroradiometer-Normalized Difference Vegetation Index. The impact of climate change and human activity on actual ET were separated and quantified. The results demonstrated that: (1) the annual pattern of actual ET per pixel exhibited the highest values for arable land (average 362.4 mm/a/pixel), followed by forest land and grassland (average of 159.6 and 142.8 mm/a/pixel, respectively). Significant increasing linear trends (p < 0.05) of 3.2 and 1.8 mm/a were detected in the arable land and forest land time series, respectively; (2) precipitation was the most significant of the selected climate factors (precipitation, average temperature, sunshine duration, and wind speed) for all ecosystems. The second most significant was wind speed; (3) human activity caused 89%, 98%, and 80% of the changes in actual ET of forest, grass, and arable land, respectively, while climate change caused 11%, 2%, and 20% of the changes in actual ET, in the Aksu River Basin during 2000–2015.