Suppressing effects of MMPs inhibitor GM6001, MMP-2/9 inhibitor Ⅰ ,MMP-2/9 inhibitor Ⅱ on migration of human lens epithelial cells / 中华实验眼科杂志

ABSTRACT

Background The primary pathologic mechanism of posterior capsular opacification (PCO) is proliferation,migration and epithelial-mesenchymal transformation of residuary lens epithelial cells (LECs) following cataract surgery.Matrix metalloproteinases (MMPs) play a role during the migration of LECs.Researches showed that GM6001,a broad inhibitor of MMPs,can arrest the migration of LECs,but as specific inhibitors of MMPs,the efficacy and safety of MMP-2/9 inhibitor Ⅰ and Ⅱ on LECs migration remain unclear.Objective This study was to determine and compare the inhibitory efficacy among GM6001,MMP-2/9 inhibitor Ⅰ and Ⅱ on human LECs and search the clinical medication to prevent PCO.Methods Human LECs were cultured and passaged in vitro,and the cells of 3-4 generation were incubated in 6-well plates.Then the cells of 70％ confluent monolayer were cultured in DMEM without fetal bovine serum for 12 hours.GM6001,MMP-2/9 inhibitor Ⅰ and Ⅱ at different concentrations (0.25,0.50,1.00,2.00,4.00,8.00,16.00,32.00,64.00,128.00 μmol/L) were added into the culture medium for 24 hours separately,and regularly cultured cells served as the control group.A bare area was made by a 200 μl sterile spear on the cell layer,and the migrated distance and inhibitory rate were calculated.The second or third generation of cells were incubated in 96-well plates at a density of 5×105/ml (200 μl/well).GM6001 (128.00 μmol/L),MMP-2/9 inhibitor Ⅰ (64.00 μmol/L) and Ⅱ (32.00 μmol/L) were added into the culture medium for 24 hours,and the cell viability was assayed by using MTT assay.Results Cultured cells grew well with irregular arrangement and presented the polygon in shape.The migrated distance was gradually reduced as the increase of concentrations of GM6001,MMP-2/9 inhibitor Ⅰ and Ⅱ,showing significant differences among the various concentration groups (GM6001:F=248.647,P＜0.05;MMP-2/9 inhibitor Ⅰ:F=357.125,P＜0.05;MP2/9 inhibitor Ⅱ:F=396.374,P＜ 0.05).The cell migrated distance in the control group was set to 1,the relative migrated distances were 0.478 ± 0.091,0.294±0.088 and 0.191 ±0.081 in the GM6001 group,MMP-2/9 inhibitor Ⅰ group and MMP-2/9 inhibitor Ⅱ group at the concentrations of 32.00 μmol/L,respectively,showing a significant difference among the groups (F =116.031,P＜0.01),and cell migrated distance was obviously shorter in the MMP-2/9 inhibitor Ⅱ group than that in the GM6001 group or MMP-2/9 inhibitor Ⅰ group (all at P＜0.01).The A values were 0.607±0.016,0.567±0.015,0.583±0.010 and 0.595 ±0.0138 in the control group,GM6001 group (128.00 μmol/L),MMP-2/9 inhibitor Ⅰ group (64.00 μmol/L) and MMP-2/9 inhibitor Ⅱ group (32.00 μmol/L),respectively,without significant difference among the groups (F=1.403,P＞0.05).Conclusions GM6001,MMP-2/9 inhibitor Ⅰ and Ⅱ reduce the mobility of human LECs effectively but do not affect the viability of the cells in vitro.MMP-2/9 inhibitor Ⅱ appears to be most dominant in inhibiting migration of human LECs.