Due to the high surface-to-volume ratio, micro/nano-electromechanical systems (MEMS/NEMS) undergo severe wear during the relative sliding. Graphene, possessing excellent mechanical, physical, and chemical properties, can achieve an ultralow friction and wear state, making it highly promising for significantly minimizing friction and wear in MEMS/NEMS. However, graphene films used in MEMS/NEMS are typically subjected to thermal annealing pretreatment during the fabrication process. To maintain optimal performance, it is particularly necessary to investigate the evolution of graphene tribological properties after high-temperature annealing. In this article, by performing nanoscale atomic force microscopy (AFM) measurements on mechanically exfoliated graphene, we reveal that the friction force on graphene decreases slightly upon annealing to approximately 200 °C, then gradually increases before rising rapidly once the annealing temperature exceeds 500 °C. Raman spectroscopy identifies that the changes in friction result from the annealing-induced compressive stress accumulation and defects creation. Our results provide deep insights for the application of graphene in sliding MEMS/NEMS. Graphical Abstract