Bearings play a key role in rolling mills, and the uniformity of their lubricant film directly affects the degree of wear of bearings and the safety of equipment. Due to long-term stress, the lubricant film inside the bearing is not uniformly distributed, resulting in uneven wear between the journal and the shaft tile, which increases the potential safety hazards in production. Traditional disassembly inspection methods are complex and time-consuming. Ultrasonic nondestructive testing technology, which has the advantages of nondestructive and adaptable, has become an effective means of assessing the thickness of the oil film in bearings. In this study, an experimental platform for calibrating the lubricant film thickness in bearings was constructed for the first time, and the acoustic characteristics of different thicknesses of the oil film were measured using ultrasonic detection equipment to verify the accuracy of the simulation process. The experimental results show that after discrete Fourier transform processing, the main features of the frequency channels of the reflected acoustic signals of different thicknesses of the oil film are consistent with the finite element simulation results, and the errors of the oil film thicknesses calculated from the reflection coefficients are within 10% of the set thicknesses, and the measurement ranges cover from 5 μm to 250 μm. Therefore, the above method can realize the accurate measurement of the thicknesses of the oil film in bearings.