Grasslands are globally important for providing essential ecosystem services and maintaining ecological security. Monitoring and assessing grassland vulnerability are critical for developing long-term grassland management policies and strategies. The grassland vulnerability on the Qinghai-Tibet Plateau (QTP) is considered high, but its spatial and temporal variations in response to human activities and climate change are not well understood. In this study, a comprehensive grassland vulnerability index (GVI), which includes natural factors (VNF), environmental disturbances (VED), and socioeconomic impacts (VSI), was developed by using the analytic hierarchy process (AHP), principal component analysis (PCA), and environmental vulnerability distance index (EVDI). Our results showed that the spatial distribution of GVI had obvious heterogeneity, decreasing from northwest to southeast; the regions with serious and extreme vulnerability were mainly concentrated in the north-western alpine steppe and desert steppe. From 2000 to 2018, GVI decreased from 0.61 in 2000 to 0.60 in 2010 and then to 0.59 in 2018, demonstrating a healthy tendency. The normalized difference vegetation index (NDVI), land desertification, and population were the factors that had the most significant impact on VNF, VED, and VSI, respectively. The global Moran’s I index of grassland vulnerability was greater than 0, with a significant positive spatial correlation. The number of High-High and Low-Low units decreased, indicating that the High-High and Low-Low cluster regions tended to be discrete. Moreover, our results suggest that understanding the variations in grassland vulnerability on the QTP is important for regional sustainable development in the context of intensified climate change and human disturbances.