The Achilles tendon enthesis (ATE) anchors the Achilles tendon into the calcaneus through fibrocartilaginous tissue. The latter is enriched in type II collagen and proteoglycans (PGs), both of which give the enthesis its capacity to withstand compressive stress. Because unloading and reloading induce remodeling of the ATE fibrocartilage (Camy et al., 2022), chronic changes in the mechanical load could modify the mechanical response under compressive stress. Therefore, we investigated the ATE fatigue behavior in mice, under cyclic compressive loading, after 14 days of hindlimb suspension and 6 days of reloading. In addition, we performed a qualitative histological study of PGs in ATE fibrocartilage. The mechanical behavior of ATE was impaired in unloaded mice. A significant loss of 27 % in Δd (difference between the maximum and minimum displacements) was observed at the end of the test. In addition, the hysteresis area decreased by approximately 27 % and the stiffness increased by over 45 %. The increased stiffness and loss of viscosity were thrice and almost twice those of the control, respectively. In the reloaded entheses, where the loss of Δd was not significant, we found a significant 28 % decrease in the hysteresis area and a 26 % increase in stiffness, both of which were higher regarding the control condition. These load-dependent changes in the mechanical response seem partly related to changes in PGs in the uncalficied part of the ATE. These findings highlight the importance of managing compressive loading on ATE when performing prophylactic and rehabilitation exercises.