A display substrate and a display device applying the same are provided. The display substrate includes a base plate and a display structure. The display structure is disposed on the base plate and includes first region. The first region includes a first sub-pixel, a second sub-pixel and two third sub-pixels. One of the two third sub-pixels has a first light emitting region having a first end point and a second end point. The other one of the two third sub-pixels has a second light emitting region having a third end point and a fourth end point. The first sub-pixel has a third light emitting region and the second sub-pixel has a fourth light emitting region. The third light emitting region and the fourth light emitting region are inside a quadrilateral region enclosed by the first, second, third and fourth end points.