Rainfall and groundwater level fluctuation lead to frequent rock slope instability accidents. In order to analyze the influence of rock mass saturation on rock slope, the relationship between saturation and uniaxial compressive strength is introduced into the shear strength of rock mass. A kinematic LA method is proposed to analyze the three-dimensional (3D) stability of rock slopes under different rock mass saturations. A hybrid optimization method is simultaneously employed to find the critical solution of the stability factor and the critical failure surface. The proposed method is confirmed to be accurate and effective through comparisons with previous solutions. The results study show that rock mass saturation significantly influences slope stability, with a sharp decline in stability observed at saturation levels below 0.6, and a more gradual stabilization beyond this point. The slope stability factor decreases significantly with the increase of the constraint width of the failure mechanism, indicating the importance of the 3D effect in slope stability analysis. The results provide practical insights for engineering applications.