The invention discloses a rolling bearing stiffness testing experiment device and an experiment method thereof. The experiment device comprises a hydraulic initial pre-tightening module, a piezoelectric actuator pre-tightening force adjustment module and a frequency response function testing module. The hydraulic initial pre-tightening module comprises a hydraulic system, a hydraulic cylinder, a tension sensor, a tension display and a spindle; the piezoelectric actuator pre-tightening force adjustment module comprises a plurality of first pressure sensors, a plurality of piezoelectric actuators, an actuator hole seat, a rolling bearing, an end cap and a bearing seat; the frequency response function testing module comprises a plurality of acceleration sensors, a plurality of second pressure sensors, a plurality of vibration exciters, a data acquisition instrument, a computer and a signal generator. According to the experiment method, a mode of implementing combined pre-tightening to the rolling bearing is adopted, and stiffness of the rolling bearing is calculated in the frequency response function testing module. By the rolling bearing stiffness testing experiment device, dynamic test on the stiffness of the rolling bearing is realized through accurate adjustment of pre-tightening force of the rolling bearing.