The invention relates to photo-crosslinking sericin protein hydrogel and a preparation method and application thereof. The preparation method comprises the steps that methyl propenyl modified sericin protein is prepared into a solution to be mixed with a photoinitiator (Irgacure 2959) solution, and the sericin protein hydrogel is obtained through ultraviolet irradiation. The obtained sericin protein hydrogel has the good biodegradability, autofluorescence characteristic and injectable property, and wounds caused by implanting or taking out biomaterials can be effectively reduced. Most importantly, the sericin protein hydrogel has good biocompatibility both in vivo and in vitro, and adhesion of carried cells can be effectively promoted. Serving as 2D and 3D cytoskeleton, the sericin protein hydrogel can effectively promote proliferation of the carried cells and promote the carried cells to form functional tissue under the condition of being free of serum support; serving as wound accessories, the sericin protein hydrogel can repair skin wounds and can serve as tissue engineering and regenerative medicine biological materials.