Abstract: A light-emitting device and a manufacturing method thereof, a display substrate and a display apparatus are provided in the embodiments of the present disclosure. The light-emitting device includes: a first electrode disposed on a base substrate; a quantum dot light-emitting layer disposed at a side of the first electrode away from the base substrate; and an electron transport layer disposed at a side of the quantum dot light-emitting layer close to or away from the first electrode, the electron transport layer including a first transport sub-layer and a second transport sub-layer which are arranged in layer configuration, a material of the first transport sub-layer and a material of the second transport sub-layer both including n-type metal oxides, and an oxygen content of the first transport sub-layer being less than an oxygen content of the second transport sub-layer.