Study On The Efficacy And Mechanism Of Luteolin In The Prevention And Treatment Of Proliferative Vitreoretinopathy

Objective: To explore the inhibitory effect of luteolin on TGF-β2-induced epithelial mesenchymal transition of ARPE-19,and the preventive and therapeutic effects and related mechanisms of proliferative vitreoretinopathy in vivo,providing a new scheme for the clinical treatment of PVR.METHODS: Human-derived retinal pigment epithelial cells,ARPE-19,and 6-8week healthy male C57BL/6J mice were used as the primary subjects for this experiment.In vitro:ARPE-19 cells were treated with different concentrations of luteolin with or without the presence of TGF-β2(10ng/ml).(1)CCK8 assays and flow cytometry were performed to assess the cellular activity of different concentrations of luteolin treatment and the effect on cycle distribution.;(2)Scratch test and Transwell migration assay for horizontal and vertical migration of cells;(3)Collagen gel contraction assay was performed to assess cell contraction;(4)Modelling of immunofluorescence and western blotting assays to detect EMT and the effect of luteolin.In vivo:To establish a mouse PVR model.(1)Retinas were extracted on 28 days and Western blot assays were performed to detect the expression of mesenchymal proteins Vimentin,α-SMA and p-ERK1/2.(2)Eyes were dewaxed and filmed and subjected to HE staining and immunofluorescence to detect the production of preretinal membranes and the severity of PVR.RESULTS: In vivo studies,we found that luteolin inhibits ARPE-19 cell proliferation in a time and concentration dependent manner within a certain concentration range;12.5 μM and 25 μM luteolin can significantly inhibit the horizontal and vertical migration ability of ARPE-19 cells;12.5 μM and 25 μM luteolin can inhibit the contraction of collagen gel promoted by TGF-β2;TGF-β2 was capable of successfully inducing EMT of ARPE-19,while 12.5μM and 25 μM luteolin can effectively inhibit the expression of mesenchymal proteins in ARPE-19 cells induced by TGF-β2.In vivo studies have found that luteolin can inhibit the increase of mesenchymal protein in the eyes of animals subjected to modeling treatment,and there is a downregulation of p-ERK1/2 protein expression during this process.CONCLUSION: Luteolin can inhibit the proliferation,migration,contraction,and TGF-β2 induced epithelial mesenchymal transformation of ARPE-19 cells in vitro.In vivo,it effectively slow down the occurrence and development of PVR by inhibiting the expression of ERK signaling pathway.