OBJECTIVE: To explore the inhibitory effect of luteolin on glaucoma-related cytokines and trabecular meshwork fibrosis, and to provide new ideas for clinical treatment of glaucoma. STUDY DESIGN: Human primary trabecular meshwork cells were cultured. Dexamethasone was applied to induce human trabecular meshwork cells to fibrosis. After luteolin intervention, transforming growth factor-beta 2 (TGF-β2), SFRP1 cytokines, and FN, COL-IV, LN, and other extracellular matrix protein expression levels were all measured to evaluate the inhibitory effect of luteolin on glaucoma-related cytokines and trabecular meshwork fibrosis. TGF-β2 overexpression adenovirus vector was used to make a high intraocular pressure mouse model, in which the luteolin intervention was explored. RESULTS: Dexamethasone can increase the expression levels of extracellular matrix proteins such as FN, COL-IV, and LN, as well as cytokines such as TGF-β2 and SFRP1 in human trabecular meshwork cells. After luteolin intervention, the expression levels of the extracellular matrix protein and glaucoma-related cytokines were significantly reduced. Luteolin can significantly reduce the intraocular pressure in mice of a high intraocular pressure model established by adenovirus vector with TGF-β2 overexpression. CONCLUSION: Luteolin can inhibit trabecular meshwork fibrosis and reduce the expression of glaucoma-related cytokines. Luteolin has the effect of reducing intraocular pressure in animal models of high intraocular pressure. This study provides an experimental basis for the treatment of glaucoma. © 2021 Science Printers and Publishers Inc.. All rights reserved.