In this paper, targeting to understand the underlying explainable factors behind observations and modeling the conditional generation process on these factors, we propose a new task, disentanglement of diffusion probabilistic models (DPMs), to take advantage of the remarkable modeling ability of DPMs. To tackle this task, we further devise an unsupervised approach named DisDiff. For the first time, we achieve disentangled representation learning in the framework of diffusion probabilistic models. Given a pre-trained DPM, DisDiff can automatically discover the inherent factors behind the image data and disentangle the gradient fields of DPM into sub-gradient fields, each conditioned on the representation of each discovered factor. We propose a novel Disentangling Loss for DisDiff to facilitate the disentanglement of the representation and sub-gradients. The extensive experiments on synthetic and real-world datasets demonstrate the effectiveness of DisDiff.