Disclosed are a display substrate, a preparation method thereof, and a display apparatus. The display substrate includes: a driving substrate, a first electrode, an auxiliary electrode, a pixel definition layer and a hole injection layer. The driving substrate includes a driving circuit and a dielectric layer covering the driving circuit; the first electrode and the auxiliary electrode are on a side of the dielectric layer of the driving substrate away from the driving circuit; the auxiliary electrode at least partially surrounds the first electrode; the pixel definition layer is on a side of the first electrode and the auxiliary electrode away from the driving substrate; the pixel definition layer includes a pixel opening, and the first electrode is at least partially exposed through the pixel opening and is electrically connected to the driving circuit; and the hole injection layer is in the pixel opening and stacked with the first electrode.