The ecological environment is important for the natural disaster prevention of human society. The monitoring of ecological environment quality has far-reaching practical significance for the functional construction of ecosystem services and policy coordination. Based on Landsat 8 operational land image (OLI)/thermal infrared sensor (TIRS) remote sensing image data, this study selected the normalized vegetation (NDVI), tasseled cap transformation humidity (WI), bare soil (SI), construction index (NDSI), and land surface temperature (LST) indexes from the aspects of greenness, humidity, dryness, and heat. Using spatial principal component analysis (SPCA) and the remote sensing ecological index (RSEI) analyzed the spatial differentiation characteristics and influencing factors of the original remote sensing ecological index (RSEI0). The results showed that: (1) the overall RSEI average value of the Qinling-Daba Mountains in 2017 was 0.61, and the ecological environment quality was at a “Good” level. Greenness contributed the most to the comprehensive index of the area, and vegetation distribution had a significant impact on the ecological environment quality of the study area. Heat is a secondary impact, and it has an inhibitory effect on habitat quality; (2) the overall distribution of regional ecological environment quality was quite different, with the ecological environment quality level showing a decreasing trend from low to high altitude; RSEI0 spatial heterogeneity at the optimal scale of 2 km was the largest, and the nugget effect was 88% which indicated a high degree of spatial variability, mainly affected by structural factors; (3) Slope, relief amplitude, elevation, the proportion of high-vegetation area, proportion of construction land area, and average population density significantly impact the spatial differentiation of RSEI0. The explanatory powers of slope and relief amplitude were 56.1% and 65.3%, respectively, which were the main factors affecting the spatial differentiation of the ecological environment quality in high undulation. The results can provide important scientific support for ecological environment construction and ecological restoration in the study area.