Suppressing effects of MMPs inhibitor GM6001, MMP-2/9 inhibitorI, MMP-2/9 inhibitorII on migration of human lens epithelial cells

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 inhibitorⅠand Ⅱ on LECs migration remain unclear. Objective This study was to determine and compare the inhibitory efficacy among GM6001, MMP-2/9 inhibitorⅠand 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 inhibitorⅠand 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 inhibitorⅠ(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 inhibitorⅠand Ⅱ, showing significant differences among the various concentration groups (GM6001: F=248. 647, P 0. 05). Conclusions GM6001, MMP-2/9 inhibitorⅠ and Ⅱ reduce the mobility of human LECs effectively but do not affect the viability of the cells in vitro. MMP-2/9 inhibitorⅡappears to be most dominant in inhibiting migration of human LECs. Key words: Matrix metalloproteinase/inhibitor; Epithelial cells/eye; Lens/ophthalmology; Migration/drug effect; Posterior capsule opacification