A key challenge to the sustainability and security of grassland capacity is the protection of water-related ecosystem services (WESs). With the change of land use, the supply of aquatic ecosystem services has changed, and the grassland-carrying capacity has been affected. However, the correlation mechanism between WESs and the grassland-carrying capacity is not clear. In this study, we used the InVEST(Integrated Valuation of Ecosystem Services and Tradeoffs) model to evaluate the impact of land-use change on WESs, and made a tradeoff analysis between WESs and grassland-carrying capacity. Considering that the Heihe River Basin (HRB) was an important grassland vegetation zone, which was a milestone for the development of animal husbandry in China, HRB was taken as a case. The main findings are as follows: (1) the spatial distribution of WESs shows the dissimilation rule, the upper reaches are the main water yield area, the soil retention is weakening in the middle and lower reaches, and the pollution has further increased in the middle and upper reaches. (2) The carrying capacity of animal husbandry decreased in the upper reaches, increased in Shandan County and Zhangye City in the middle reaches, and decreased sharply in other regions. (3) There was a positive correlation between the livestock-carrying capacity and nitrogen export in 2018, which was increasing. As the change of land use has changed the evapotranspiration structure, WESs have undergone irreversible changes. Meanwhile, the development of large-scale irrigated farmland and human activities would be the source of a further intensification of regional soil erosion and water pollution. Therefore, it is necessary to trade off the WESs and animal husbandry under land-use change. This paper revealed how WESs changed from 2000 to 2018, the characteristics of the changes in the spatial and temporal distribution, and the carrying capacity. It aims to provide a scientific basis for coordinating the contradiction between grassland and livestock resources, improving the regional ecological security situation, and carrying out ecosystem management.