Objective To observe the effect of bone forming protein 4(BMP4)on the proliferation and migration of human retinal pigment epithelium(RPE)cells under oxidative stress,and to preliminarily explore its effect on epithelial-mesenchymal transition(EMT)of RPE cells.Methods Human RPE cells cultured in vitro were divided into normal group,pure 4-hydroxynonenal(HNE)group(4-HNE group),4-HNE+NC group and 4-HNE+small interfering BMP(siBMP4)group.The effect of 4-HNE on the proliferation of RPE cells was detected by thiazole blue colorimetry.The effects of 4-HNE and BMP4 on cell migration were determined by cell scratch test.The expression of BMP4 was detected by immunofluorescence staining,Western blot and real-time quantitative polymerase chain reaction.The transfection efficiency of siBMP4 was observed by fluorescence microscopy.Mitochondrial reactive oxygen species(MitoSOX)were detected by flow cytometry.The expression of EMT markers E-cadherin and Fibronection were detected by immunofluorescence assay.t-test was used for comparison between the two groups,and one-way analysis of variance was used for comparison between the three groups.Results Compared with normal group,cell proliferation and migration ability of 4-HNE group were significantly enhanced,with statistical significance(t=21.619,24.469;P＜0.05).The expression of BMP4 in cells was significantly increased,and the difference was statistically significant(t=19.441,P＜0.05).The relative expression levels of BMP4 mRNA and protein were also significantly increased,with statistical significance(t=26.163,37.163;P＜0.05).After transfection with siBMP4 for 24 h,the transfection efficiency of BMP4 in RPE cells was＞90％.Compared with 4-HNE group and 4-HNE+NC group,the relative expression levels of BMP4 protein(F=27.241),mRNA(F=36.943),cell mobility(F=46.723)and MitoSOX expression levels(F=39.721)in normal group and 4-HNE+siBMP4 group were significantly decreased.The differences were statistically significant(P＜0.05).The epithelial marker E-cadherin increased significantly,while the mesenchymal marker Fibronection decreased significantly,with statistical significance(F=51.722,45.153;P＜0.05).Conclusions BMP4 inhibits RPE proliferation and migration under oxidative stress.BMP4 is involved in inducing EMT in RPE cells.
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