A display substrate, a method for fabricating the same, and a display panel are provided. The display substrate includes: a substrate, and a first conductive layer, at least two insulation layers, and a second conductive layer, the second conductive layer being electrically connected with the first conductive layer through via-holes, and the at least two insulation layers including a first insulation layer in contact with the first conductive layer, wherein the display substrate further includes an assisting alignment structure on the surface of the first insulation layer, and the orthographic projection of the assisting alignment structure surrounds at least part of the edge of the orthographic projection of the first via-hole in the first insulation layer on the substrate, so that the orthographic projection of the first via-hole on the first conductive layer lies within the pattern of the first conductive layer.