ABSTRACT
AIM: To investigate the effect and mechanism of matrix metalloproteinase(MMPs)inhibitor AG3340 on the migration and invasion ability of retinal pigment epithelial cells-19(ARPE-19)cultured in high glucose(CHG). METHODS: ARPE-19 cells cultured in vitro were divided into four groups: Control group, the glucose at the concentration of 5.6mmol/L in DMEM/F12 medium; HG group, the glucose at the concentration of 30mmol/L was cultured with DMEM/F12 medium; HG+AG3340 group, the cells were pretreated with AG3340 for 12h, and then cultured in DMEM/F12 medium containing 30mmol/L glucose; The mannitol(MA)group, cultured with DMEM/F12 medium of 5.6mmol/L glucose and 24.4mmol/L mannitol, which used as hypertonic control group. The migration ability of cells was detected by wound healing assay, the invasion ability of cells was detected by Transwell assay, and the relative expression levels of MMP-9, MMP-2, fibronectin and collagen Ⅳ were detected by Western blot.RESULTS: The results of wound healing assay showed that compared with the Control group, the cell migration rate of scratching after 24h and 48h in the HG group was significantly increased(all P<0.001).After pretreated by AG3340, the cell migration rate was significantly lower than that in the HG group(all P<0.01).Transwell assay showed that compared with the Control group, the number of cell invasion in the HG group was significantly higher than that in the Control group(all P<0.001). After pretreated by AG3340, the number of cell invasion was decreased than the HG group(all P<0.01). Western blot results showed that compared with the Control group, the relative expression levels of MMP-9 and MMP-2 of the cells in the HG group were increased, and the relative expression levels of Fibronectin and Collagen Ⅳ were decreased(all P<0.001). Compared with the HG group, the relative expression levels of MMP-9 and MMP-2 protein in AG3340 pretreatment group were decreased, and the relative expression levels of Fibronectin and Collagen Ⅳ were increased(all P<0.05). CONCLUSION: High glucose induced ARPE-19 cells with enhanced migration and invasion ability, and AG3340 partially reversed this effect, which was related to the inhibition of MMP-9 and MMP-2 expression and the stability of extra-cellular matrix components.
ABSTRACT
AIM:To observe the pathological changes of the lower segment of nasolacrimal duct mucosa in rabbits at different stages after retrograde lachrymal dilated drainage tube implantation. METHODS:Totally 14 New Zealand rabbits were used in the present study. One side of nasolacrimal duct was obstructed to produce an experimental model and operated the reverse implantation of nasolacrimal duct intubation. Histological changes of the lower segment of nasolacrimal duct mucosa were observed by routine light microscope at 2, 4, 6, 8, 10, 12 and 14wk after the operation. RESULTS: Compared with the control side, the group of 2 and 4wk after surgery presented the inflammatory cytokine. The group of 12wk after the operation presented isolated granuloma. Group 12 and 14wk presented scattered granuloma. The size of the granulomas was smaller and the density of epithelioid cell and fibroblast were lower in group 12wk than those in group 14wk by HE and Masson trichrome stain. CONCLUSION:Recurrent Silicone Tube is used to treat nasolacrymal duct obstruction. Nasolacrimal duct can be narrowed and blocked again by granuloma, progressive fibrosis and adhesion of surrounding tissues when tube is in the duct more than 12wk.