The phenomenon of the oxidation and aging of asphalt binders affects the strength and durability of asphalt mixtures in pavements. Several studies are trying to improve the resistance to this phenomenon by modifying the properties of the binders with nano-particles. One material that shows promise in this field is zinc oxide (ZnO), especially in improving ultraviolet (UV) aging resistance. Few studies have evaluated the effect of these nano-particles on the thermo-oxidative resistance of asphalt binders, and, on hot-mix asphalt (HMA), studies are even more scarce and limited. Therefore, in the present study, the resistance to thermo-oxidative aging of an HMA manufactured with an asphalt binder modified with ZnO was evaluated. An asphalt cement (AC 60–70) was initially modified with 0, 1, 3, 5, 7.5, and 10% ZnO (percentage by weight of asphalt binder; ZnO/AC in wt%), and then exposed to aging in Rolling Thin-Film Oven tests (RTFOT) and a Pressure Aging Vessel (PAV). Penetration, viscosity, and softening point tests were performed on these binders, and aging indices were calculated and evaluated. Samples of HMAs were then manufactured using these binders and designed by the Marshall method, determining the optimum asphalt binder content (OAC) and the optimum ZnO/AC ratio. Control (unmodified) and modified HMA were subjected to short-term oven aging (STOA) and long-term oven aging (LTOA) procedures. Marshall, Indirect Tensile Strength (ITS), and resilient modulus (RM) tests were performed on these mixtures. LTOA/STOA results of the parameters measured in these tests were used as aging indices. In this study, ZnO was shown to increase the thermo-oxidative aging resistance of the asphalt binder and HMA. It also contributed to an increase in the resistance under monotonic loading in the Marshall and ITS tests, and under repeated loading in RM test. Likewise, it contributed to a slightly increasing resistance to moisture damage. The best performance is achieved using ZnO/AC = 5 wt%.