Poor subgrade conditions usually induce various subgrade diseases in railways, leading to some adverse influences. An innovative technology that involves installing a prestressed reinforcement structure (PRS) that consists of steel bars and lateral pressure plates (LPP) for subgrade was introduced to improve its stress field and provide compulsive lateral deformation constraints for slope. In this study, an investigation into the dynamic acceleration responses of railway subgrade strengthened according to different PRS schemes was presented using a 1:5 scale model test, aiming to explore the effects of the axle load, the reinforcement pressure, and the loading cycles on the acceleration characteristics of the subgrade. The experimental results showed that (1) after pretension of the steel bar, prestress loss occurred due to the soil creep behavior and group anchor effect, so a moderate amount of over-tension in practices would be necessary; (2) a distinctive periodical behavior of subgrade subjected to the cyclic loads was observed, the horizontal accelerations were generally less than the vertical accelerations at the same measurement heights, and the vibration energy attenuated gradually from the shoulder to the toe along the slope; (3) in the short-term tests, the peak accelerations at all measurement points had a linear correlation with the axle load, and oppositely, it showed an approximately linear decrease with the increasing reinforcement pressure; And (4) in the long-term tests, to simulate the heavy haul wagon with a 35 t axle load, the variation in the effective acceleration with loading cycles under reinforcement pressure 100 kPa initially exhibited a decrease and subsequently tended to be stable, which is apparently less than that without reinforcement pressure. Consequently, it was demonstrated that the PRS itself and increasing reinforcement pressure can effectively mitigate the subgrade vibration, and provide an appropriate alternative to improve the dynamic performance of railway subgrade under the moving train loads.