Transforming growth factor-beta2 (TGF-beta 2) induces Endothelial-Mesenchymal transition (EndoMT) and autophagy in a variety of cells. Previous studies have indicated that activation of autophagy might decrease TGF-beta 2 induced EndoMT. However, the precise role remains unclear. In the present study, we found that TGF-beta 2 could induce EndoMT and autophagy in human retinal microvascular endothelial cells (hRMECs). Activation of autophagy by Rapamycin or Trehalose could reduce the expression of Snail, demonstrating a role of autophagy in regulating Snail production both by transcriptional and post transcriptional mechanism. Co-immunoprecipitation (ColP) demonstrated that LC3 coimmunoprecipitated with Smad3 and western blot showed that autophagy inducers, Rapamycin and Trehalose, could decrease the phosphorylation level of Smad3. Therefore, our results demonstrate that autophagy counteracts the EndoMT process triggered by TGF-beta 2 by decreasing the phosphorylation level of Smad3. (C) 2017 Elsevier Inc. All rights reserved.