In rheological testing, different test regimes can result in large differences in the rheological curves, and the pre-shear procedure is one of the most important parts of the test. In order to investigate the effect of the pre-shear regime on the rheological properties of cement pastes, an Anton Paar 102 rotary rheometer was used for the test. Two different pre-shear regimes (RF and CP pre-shear), three different pre-shear duration times (50 s, 100 s, 150 s), and four different pre-shear rates (25 s-1, 50 s-1, 100 s-1) are carried out for the test. The results showed that with the increase of pre-shear duration or rate, the four rheological parameters (plastic viscosity, yield stress, rheological index, and thixotropy) of the pastes decrease in different degrees. RF pre-shear was more influenced by shear time, while CP pre-shear was influenced by shear time and rate. The pre-shear regimes led to different changes in the flocculation structure of the cement paste, which had a great influence on rheological parameters ultimately. Based on the improved Krieger-Dougherty model, the effective solid volume fraction of cement paste was calculated. Analysis shows that the phenomenon of "deflocculation" and "re-flocculation" occurs when the cement paste is subjected to shear action, thus, the pre-shear regimes destroy the structure of the pastes. With the increase of pre-shear duration or rate, the effective volume fraction decreased, which indicated that the initial flocculation structure was broken. The greater the damage degree of pre-shear, the slower the establishment rate of the internal structure of the pastes in dynamic shear, resulting in lower rheological parameters.