In this paper, the discrepancy between the calculated and actual fatigue life distribution trend of the axles of railway vehicle structures is corrected. The calculations were made using the traditional fixed-axle model in which the wheelset rotation is neglected in random vibration analysis. A correction method is proposed, which is based on the original methods of fatigue life analysis of the axle structure of railway vehicles (i.e. the pseudo-excitation method and finite element model) and according to the characteristics of the symmetrical axle structure and the actual rotational operation state of the railway vehicle wheelset. Taking a locomotive system as an example, the minimum fatigue life calculated by the fixed-axle model is only 0.81 million km. In contrast, the minimum fatigue life calculated by the proposed method is 5.85 million km, which is consistent with the actual fatigue life of the axle. The position of the minimum fatigue life is also consistent with the actual position. The reliability of the proposed method for calculating the random fatigue life of the axle structure is illustrated. The influence and contribution of the prestress generated by axle press-fitting and vehicle serving weight of the railway vehicle acting on the axle on the axle fatigue life were studied. The results show that the prestress has a different influence on the fatigue life reduction of the wheelset. The minimum fatigue life reduction can reach 16.68% when all the prestresses are considered. Therefore, the influence of prestress should be considered in the design of railway vehicle axle structures. The idea of considering prestress should be proposed when revising axle standards.