The application discloses a nerve regeneration guide conduit for promoting nerve cell differentiation and a preparation method and application thereof. The nerve regeneration guide conduit can realize stable and long-acting release of bioactive factors, effectively reduce the risk of tissue adhesion, efficiently induce nerve cell differentiation, promote axon growth and function recovery, and provides a novel and effective solution for nerve regeneration and repair. In addition, the nerve regeneration guide conduit has excellent conductivity, biocompatibility, mechanical adaptability and the like, and therefore has a wide application prospect in the field of nerve regeneration and repair.