Railway wheels are usually attached to axles by press-fitting; therefore, the mechanical processes taking place during operation can result in failure, with fatal consequences for the axle seats. This manuscript describes the effect of laser hardening on the residual stress state, microstructural parameters (lattice defects—dislocations, crystallites, microstrains, etc.), and mechanical properties of laser-hardened EA1N steel railway axles under fatigue life conditions. Differences were found between ground, single-track, and multi-track hardened surfaces. Tensile residual stresses, low dislocation densities and hardnesses, and different microstructures (tempered cubic martensite) were found at the overlapped tracks and at the boundary of the heat-affected zone and bulk surface compared with the hardened zone. As a result, the surface treatment of axle seats by laser hardening improved the fatigue failure resistance compared with untreated seats. Optimal properties of the integrity of the axle seat surface were achieved, including fatigue resistance, which seems to be positively influenced mainly by sufficient hardness and the appropriate microstructure. The influence of the other investigated parameters was not evident, and was reduced by the presence of fretting corrosion and press-fitting.