Abstract: Objective:To investigate the inhibitory effect of doxycycline (Doxy) on Transforming Growth Factor-β1 (TGF-β1)-induced human corneal stromal cell fibrosis. Methods:Human corneal stromal cells were extracted from corneal stromal lens after femtosecond laser-assisted small incision lenticule extraction (SMILE), and the corneal fibrosis model was established in vitro by inducing human corneal stromal cells with TGF-β1. Control group, TGF-β1 group and Doxy group were set up. The proliferation ability of each group was detected by cell counting kit-8 (CCK-8) method, the proliferation ability was detected by scratch test, and the content of hydroxyproline (HYP), a key factor in collagen synthesis, was determined by enzyme-related immunosorbent assay (ELISA). Reverse transcription-quantitative PCR (RT-qPCR) was used to detect the mRNA expression of the key factors of fibrosis Thy-1 and Vimentin, and western blotting was used to detect the protein expression levels of Thy-1 and Vimentin. Results:Compared with the control group, TGF-β1 group could promote the proliferation and migration of human corneal stromal cells, and increase the content of HYP, the expression of Thy-1 and Vimentin (all P＜0.05). Compared with the TGF-β1 group, the abilities of proliferation, migration and collagen synthesis were significantly decreased in the Doxy group, and the expression of Thy-1 and Vimentin was decreased (all P< 0.05). Conclusion:Doxy can inhibit the proliferation, migration and collagen synthesis of human corneal stromal cells induced by TGF-β1, and down-regulate the expression levels of Thy-1 and Vinmentin, thereby inhibiting corneal fibrosis.