The invention discloses self-healing anti-freezing conductive fibroin hydrogel and a preparation method thereof, and belongs to the field of flexible electronic materials. The preparation method comprises the following steps: firstly, modifying pyrrole by adopting dopamine, polymerizing in an acid environment to obtain dopamine modified polypyrrole, then taking MAX powder as a raw material, etching under hydrofluoric acid and hydrochloric acid to obtain an MXene material, then respectively adding the dopamine modified polypyrrole and the MXene material into a polyvinyl alcohol aqueous solution, uniformly mixing, then adding a fibroin aqueous solution, further mixing, and finally, preparing the modified polypyrrole/MXene composite material. And finally, dropwise adding a mixed aqueous solution of borax and glycerol into the mixed solution to prepare the self-healing anti-freezing conductive fibroin hydrogel. The self-healing anti-freezing conductive fibroin hydrogel has the advantages of self-healing, freezing resistance, air permeability and good conductivity, overcomes the three defects of no freezing resistance, no self-healing and non-uniform conductive material distribution of common fibroin hydrogel, and has a wide application prospect in the field of flexible electronic products.