Peptides self-assembly has great potential applications in the field of biomedicine and construction of new materials for the advantages of stable structure, easy control, and good biocompatibility and biodegradability. In this paper, the progress in peptides self-assembly is systematically reviewed, including the conception, mechanism and applications, focusing on the stimuli-responsive peptides self-assembly. According to the difference of stimuli, stimuli-responsive peptides self-assembly can be classified into pH-responsive self-assembly, temperature-responsive self-assembly, solvent-responsive self-assembly, light-responsive self-assembly, ultrasonic responsive self-assembly, and ion-responsive self-assembly. The applications of peptides self-assembly in drug release carriers, the repair of spinal cord injury, biomimetic enzyme catalysis, and biological template are listed in detail. Finally, the problems of peptides self-assembly research, such as difficult control of the external factors that affect the structure of assembly and the low degree of overlap between peptides self-assembly and life sciences, are analyzed, and the future development of stimuli-responsive peptides self-assembly is prospected.