In view of the complicated cabin structure of the Three Gorges ship lift and the difficulty in selecting the equipment inspection route, the inspection route in the ship lift cabin was taken as the research object and the planning for the route was converted into a Traveling Salesman Problem (TSP). Through the weighted undirected graph of the inspection route and the spatial coordinates of the inspection points, a spatial structure model of the inspection points of the ship lift was built. The ant colony algorithm was applied to calculate the optimal inspection route for day shift and swing shift, respectively, via the Matlab software. The experimental results show that the optimal inspection route calculated by the ant colony algorithm meets the equipment inspection requirements of the Three Gorges ship lift.