Objectives Diabetic nephropathy (DN), a severe microvascular complication of diabetes mellitus, is a leading cause of end-stage renal disease. Crocin (CRO), an active ingredient extracted from Crocus sativus and Gardenia jasminoides, has multiple bioactivities such as anti-oxidative, anti-inflammatory, anti-tumor, and anti-depressive activities. However, the potential effects and mechanisms of CRO in the treatment of DN are still unclear. Methods In this study, we aimed to assess the efficacy of CRO in treating DN using in vivo and in vitro experiments, and intensively investigate the potential therapeutic mechanisms of CRO against DN based on the inhibition of epithelial-mesenchymal transition (EMT) by inducing adenosine monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR)-mediated autophagy. Results The results showed that CRO had a therapeutic effect and anti-EMT effect in kidney of DN mice. CRO also moderated AMPK/mTOR pathway and improved autophagy in kidney of DN mice. In high glucose (HG)-induced tubular epithelial cell EMT model, CRO inhibited EMT, moderated AMPK/mTOR pathway and improved autophagy. AMPK inhibitor abolished the above effects of CRO on tubular epithelial cells. Conclusion CRO exhibited considerably therapeutic and anti-EMT effects on DN both in vivo and in vitro, these may be associated with restoring autophagy through regulating AMPK/mTOR pathway.