Alxa League of Inner Mongolia Autonomous Region is a concentrated desert distribution area in China, and the latest desertification process and its driving mechanism under the comprehensive influence of the extreme dry climate and intense human activities has attracted much attention. Landsat data, including ETM+ images obtained in 2000, TM images obtained in 2010, and OLI images obtained in 2020, were used to extract three periods of desertification land information using the classification and regression tree (CART) decision tree classification method in Alxa League. The spatio-temporal variation characteristics of desertification land were analyzed by combining the transfer matrix and barycenter migration model; the effects of climate change and human activities on regional desertification evolution were separated and recombined using the multiple regression residual analysis method and by considering the influence of non-zonal factors. The results showed that from 2000 to 2020, the overall area of desertification land in Alxa League was reduced, the desertification degree was alleviated, the desertification trend was reversed, and the desertification degree in the northern part of the region was more serious than in the southern part. The barycenter of the slight, moderate, and severe desertification land migrated to the southeast, whereas the serious desertification land’s barycenter migrated to the northwest in the period of 2000–2010; however, all of them hardly moved from 2010 to 2020. The degree of desertification reversal in the south was more significant than in the north. Regional desertification reversal was mainly influenced by the combination of human activities and climate change, and the area accounted for 61.5%; meanwhile, the localized desertification development was mainly affected by human activities and accounted for 76.8%.