A display substrate and a display device are provided. The display substrate includes a plurality of repeat units. Each of the plurality of repeat units includes one first-color sub-pixel, one second-color sub-pixel pair and one third-color sub-pixel which are arranged in a first direction, the second-color sub-pixel pair includes two second-color sub-pixels arranging in a second direction. In three sub-pixels of different colors which are adjacent to each other, orthographic projections of light-emitting layers of at least two sub-pixels adjacent to each other on the base substrate are overlapped or connected, and a spacing is disposed between at least part of the light-emitting layers of two sub-pixels which are adjacent to each other.