With the development of nanotechnology and materials science, bioinspired nanochannels appeared by mimicking the intelligent functions of biological ion channels. They have attracted a great deal of attention in recent years due to their controllable structure and tunable chemical properties. Inspired by the layered microstructure of nacre, 2D layered materials as excellent matrix material of nanochannel come into our field of vision. Bionic nanochannels based on 2D materials have the advantages of facile preparation, tunable channel size and length, easy expansion, and modification, etc. Therefore, the 2D layered nanofluid system based on bionic nanochannels from 2D layered materials has great potential in biomimetic microsensors, membrane separations, energy conversion, and so on. In this paper, we focus on the construction and application of bionic nanochannels based on 2D layer materials. First, a basic understanding of nanochannels based on 2D materials is briefly introduced, we also present the property of the 2D materials and construction strategies of bionic nanochannels. Subsequently, the application of these nanochannels in responsive channels and energy conversion is discussed. The unsolved challenges and prospects of 2D materials-based nanochannels are proposed in the end.