Qinghai Province, which is the source of three major rivers (i.e., Yangtze River, Yellow River and Lancang River) in East Asia, has experienced severe grassland degradation in past decades. The aim of this work was to analyze the impacts of climate change and human activities on grassland ecosystem at different spatial and temporal scales. For this purpose, the regression and residual analysis were used based on the data from remote sensing data and meteorological stations. The results show that the effect of climate change was much greater in the areas exhibiting vigorous vegetation growth. The grassland degradation was strongly correlated with the climate factors in the study area except Haixi Prefecture. Temporal and spatial heterogeneity in the quality of grassland were also detected, which was probably mainly because of the effects of human activities. In the 1980s, human activities and grassland vegetation growth were in equilibrium, which means the influence of human activities was in balance with that of climate change. However, in the 1990s, significant grassland degradation linked to human activities was observed, primarily in the Three-River Headwaters Region. Since the 21(st) century, this adverse trend continued in the Qinghai Lake area and near the northern provincial boundaries, opposite to what were observed in the eastern part of study. These results are consistent with the currently status of grassland degradation in Qinghai Province, which could serve as a basis for the local grassland management and restoration programs.