This study aims to evaluate the effects and mechanisms of dendrobine against thyroid-associated ophthalmopathy (TAO). Potential targets of dendrobine and TAO were selected to construct a protein-protein interaction network, elucidating the potential mechanisms of dendrobine against TAO. The potential mechanisms systematically were verified by molecular docking, transcriptomics and in vitro experiments. Orbital fibroblasts (OFs) were isolated from orbital decompression-derived fat tissue, and the effects of dendrobine were studied in transforming growth factor-beta 1 (TGF-β1)-induced fibrosis models and interleukin-1β (IL-1β)-induced inflammation models. The results showed that dendrobine inhibited the migration, fibrosis and levels of inflammatory factors in TAO OFs. Network pharmacology and transcriptomics studies revealed the mechanisms of dendrobine, while in vitro experiments confirmed it alleviates TGF-β1-induced fibrosis in TAO OFs by inhibiting AKT phosphorylation.