Fig. 4

NF-κB mediated TRIM46 expression and is involved in the effects of TRIM46 on HRCECs. a TRIM46 expression in HRCECs was detected by Western blot and qRT-PCR after treatment with HG (25 mM) and 10 μM NF-κB inhibitor PDTC at different time points. b HRCECs were treated with HG (25 mM) and treated with 10 μM NF-κB inhibitor PDTC for 24 h. Transcriptional activity of TRIM46 promoter was analyzed by the luciferase reporter gene assay. c–h HRCECs were transfected with oeTRIM46 or Vector and were administered with NF-κB inhibitor PDTC (10 μM) and HG (25 mM). Cell proliferation and cell cycle was detected with CCK-8 assay (c) and flow cytometry (d), respectively. Transmembrane electrical resistance assay (e) and FITC-dextran leak assay (f) for analyzing cell permeability were performed after the cells grew to confluence. g Western blot showing the expression levels of TRIM46, Occludin and ZO-1. h ELISA used for the detection of TNF-α, IL-1β and IL-6 levels in supernatant of cells. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001. HG, high glucose; CCK, cell counting kit; ELISA, enzyme-linked immunosorbent assay; FITC, fluorescein isothiocyanate; HRCECs, human retinal capillary endothelial cells; NF-κB, nuclear factor kappa B; PDTC, pyrrolidine dithiocarbamate; TRIM, tripartite motif