Estimating the spatiotemporal variations in natural grassland carrying capacity is crucial for maintaining the balance between grasslands and livestock. However, accurately assessing this capacity presents significant challenges due to the high costs of biomass measurement and the impact of human activities. In this study, we propose a novel method to estimate grassland carrying capacity based on potential net primary productivity (NPP), applied to the source area of the Nujiang River and Selinco Lake on the Tibetan Plateau. Initially, we utilize multisource remote sensing data—including soil, topography, and climate information—and employ the random forest regression algorithm to model potential NPP in areas where grazing is banned. The construction of the random forest model involves rigorous feature selection and hyperparameter optimization, enhancing the model’s accuracy. Next, we apply this trained model to areas with grazing, ensuring a more accurate estimation of grassland carrying capacity. Finally, we analyze the spatiotemporal variations in grassland carrying capacity. The main results showed that the model achieved a high level of precision, with a root mean square error (RMSE) of 4.89, indicating reliable predictions of grassland carrying capacity. From 2001 to 2020, the average carrying capacity was estimated at 9.44 SU/km2, demonstrating a spatial distribution that decreases from southeast to northwest. A slight overall increase in carrying capacity was observed, with 65.7% of the area exhibiting an increasing trend, suggesting that climate change has a modest positive effect on the recovery of grassland carrying capacity. Most of the grassland carrying capacity is found in areas below 5000 m in altitude, with alpine meadows and alpine meadow steppes below 4750 m being particularly suitable for grazing. Given that the overall grassland carrying capacity remains low, it is crucial to strictly control local grazing intensity to mitigate the adverse impacts of human activities. This study provides a solid scientific foundation for developing targeted grassland management and protection policies.