The change in the composite lay-up method affects the blade stiffness, which in turn affects the structural dynamic and aerodynamic characteristics, but the influence law is not yet clear. In this paper, two-way fluid–structure coupling is used to study wind turbine blades with different numbers of lay-up layers. The analysis results show the following: (1) With the increase in the number of pavement layers, the maximum deformation of the blade tip decreases, the magnitude of the decrease also decreases gradually, and the growth trend of deformation along the blade spreading direction is gradually flattened. (2) The maximum stress–strain of the root of the blade gradually decreases, the tip of the blade is the opposite of the blade, the unevenness of the distribution of the stress–strain of the surface of the blade is gradually slowed down, and the concentration area is shifted back. (3) The different numbers of pavement layers affect the turbulence intensity of the blade in the flow field space. Herein, the spatial distribution trend of turbulence intensity in the flow field of wind turbine blades with different numbers of layers is basically the same, and the trend of turbulence intensity changes is gentle when the number of layers is increased. (4) The amount of the tip vortex in the wake stream of the wind turbine decreases, and the decay rate of the center vortex increases. A reasonable lay-up scheme can improve the deformation and stress of the blade, extend the service life of the blade, promote airflow mixing, and enhance the energy conversion efficiency.