RESUMO
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.
RESUMO
?AIM:To analyze the morphological changes of macular soft drusen and drusenoid pigmental epithulium detachment ( DPED ) after subthreshold micropulse laser treatment ( SMLT) .?METHODS: Fourteen patients ( 20 affected eyes ) with soft drusen and DPED clinically confirmed from August 2016 to October 2018, were included in this study. 577 nm yellow laser of SMLT was applied for soft drusen and DPED. The changes of soft drusen and DPED in best corrected visual acuity ( BCVA ) ( LogMAR ) and height, diameter and cross-sessional area according to fundus autofluorescence and SD - OCT examinations were observed after SMLT.?RESULTS: BCVA was not significant difference after treatment of soft drusen (P=0.260), and the DPED (P=0. 736 ) than that of the baseline. Compared with the baseline values, the height and cross-sessional area of soft drusen were reduced at the 6mo after treatment ( P=0. 008, P=0.034) . Compared with the baseline values, the differences were not statistically significant in height, diameter and cross - sectional area of DPED after treatment.?CONCLUSION: BCVA was not reduced for drusen and DPED after SMLT, however, the height and cross -sessional area of soft drusen was reduced compared with those before treatment, and the differences were not statistically significant in height, diameter and cross -sectional area of DPED before and after treatment. The results indicated that SMLT was effective for soft drusen, but was not effective for short-term treatment of DPED. SMLT caused no damage to the visual acuity in treatment of soft drusen and DPED, but prospective, controlled, large sample and long-term follow-up studies should be required.