The ecological security of the Tibetan Plateau is vital for sustainable development. In recent years, biodiversity loss and ecosystem degradation caused by industrial and mining activities have attracted wide attention. However, a synthesis assessment of the impacts of industrial and mining land (IML) on the ecosystem is currently lacking. In this study, based on the land cover data and normalized differential vegetation index, we used the landscape ecological index, Theil-Sen trend analysis and equivalent value factors method to evaluate the change in IML and its ecosystem impact on the TP. The results demonstrated that the area of IML expanded by 3.3 times (228.56%) during 1990–2020, and reached 968.95 km² in 2020. Within this area, the newly added, stable, and reduced areas were 842.71, 126.26, and 168.65 km², respectively. Simultaneously, IML expansion made the landscape more fragmented during 1990–2020. The number of patches, splitting index, and landscape shape index in 2020 increased by 3.59-, 2.70-, and 1.90-fold compared to those in 1990, respectively. Furthermore, the difference in the vegetation change between the IML and its 10 km buffer zone was significant. About 77.34% of the vegetation in the IML area showed a trend of decrease, while about 76.51% of the vegetation in the buffer zone of IML showed a trend of increase. In addition, the expansion of IML also reduced the total ecological services value by USD 6969.31 million (0.66%) from 1990 to 2020. However, the lowered value was USD 8649.50 million (0.82%) in the newly added IML. This study highlights the rapid expansion of IML and reveals the ecosystem structure, ecosystem quality, and ecosystem service impact on the TP, which helps guide ecosystem protection and the sustainable development of mining.