In this study, a 3-year experiment with different grazing intensities was conducted in a temperate grassland enclosed for 5 years in Lhasa Valley grassland in order to provide scientific evidence to conservation of grassland ecological environment on the Qinghai-Tibet Plateau. Soil samples were collected and soil protozoa were cultured and identified by non-submerged culture and in vivo observation in September of 2018. The results showed that a total of 46 soil protozoa genera were identified, belonging to 36 families, 21 orders, 12 classes and 2 phyla. The main group of soil protozoa was Spirotrichea under different grazing intensities. The diversity indexes (H,E, C, M) of soil protozoa had no significant difference under different grazing intensities (P>0.05), Jaccard similarity index ranged from extremely dissimilar to moderately dissimilar. Plot B3 (ban grazing) was clustered into a single category, indicating that the spatial heterogeneity was large. TN was the main environmental factor affecting the distribution of soil protozoa, the combined effects of soil water content (SWC), rapid available potassium (RAK), EC and other environmental factors affected the species composition and distribution of soil protozoa under different grazing intensities.