Changes in the spatial pattern of land use are inextricably linked to the ecosystem environment, and the assessment of regional eco-environment quality can help provide sustainable and healthy development strategies for heritage management organizations. In this study, based on RS and GIS technologies, we quantitatively analyzed the characteristics of land use changes in the Shibing Karst World Natural Heritage site from 2000 to 2020, and introduced the regional eco-environment quality assessment index for quantitative factor analysis. The results show that: (1) The heritage site is mainly dominated by forest and shrub, with more aggregated cropland, and impervious surfaces in the buffer zone. The area of shrub has increased during the 20-year period, occupying 12.63% of the total transferred area. Cropland has been basically converted to forest, accounting for more than 60% of the total transferred area, followed by shrubs, which have been basically transferred to ecologically better land types. (2) By analyzing the attitude of motivation, forest, shrub, and water have positive values, while cropland, grassland, and impervious surfaces have negative values. Grassland has the largest absolute value of kinetic attitude, and the smallest is for water. The integrated dynamic attitude is basically maintained at 24%, showing a state of rapid and then stable land category conversion. (3) Over the past 20 years, the regional eco-environment quality index has been stable between 0.68 and 0.71, and shows a trend of rapid growth and stabilization, which is consistent with the comprehensive attitude. The conversion between cropland, forest, and shrubs is the main cause of ecological improvement and deterioration. Overall, the relevant conservation measures at the site and China’s corresponding responses to global climate change have led to a stabilization and increase in the regional ecological quality of the site. The management measures of the relevant authorities have begun to bear fruit, but further promotion of the sustainable development of the site is needed to provide a scientific model for the conservation of other karst heritage sites.