The Effect Of Matrix Metalloproteinase Inhibitor On Corneal Wound Healing Of Rabbits After LASIK

Objective:Combing the mothed of molecular biology,immunobiology and the technology of confocal microscope,To study the realition of gelatinase and corneal wound healing of rabbits after laser in situ keratomileusis LASIK.To prove whether matrix metalloproteinase inhibitor has effect on corneal wound healing.This test is to further understand the mechanism of corneal wound healing response after LASIK.Methods:1,40 rabbits were treated by LASIK surgery both eyes.Left eyes(experiment group)of each was treated with GM6001(200μg/ml)and PBS with right eyes(control group).Both group were injected the liquid once a day under the conjunctiva with the same procession.Finished 30d later.2,Counted the numbers of stroma cells and endothelium cells of rabbits before LASIK surgery using confocal microscope.The all groups of rabbits were examined at 1d,3d,7d,1m and 3m after the surgery and counted the numbers of stroma cell,strong-lighted spots and endothelium cell each days.Sacrificed the experimental rabbits at 1d,3d,7d,1m and 3m after the surgery and aquired the cornea to perform indirect enzyme-linked immuno-absordent assay (ELISA)for the quantity of gelatinase.,zymography for the activity of gelatinase and indirect immuno flurescent for the location of gelatinase.Result:1,The confocal microscope proved that the stroma cell undergone apoptosis after LASIK.The loss numbers of the stroma cells in experimental group were less than in control group at 1d,3d,7d,1m and 3m after the surgery.The differences were statistically significant (F=4.511,P＜0.05).The numbers of spots in stroma layer of the experimental group were less than the control group at 1d,3d,7d,1m and 3m after the surgery.The differences were statistically significant(F=5.001,P＜0.05).The lost numbers of endothelium cells in experimental group were the same with in control grouP at 1d,3d,7d,1m and 3m after the surgery.The differences have no statistically significant(P＞0.05).2,Indirect enzyme-linked immuno-absordent assay(ELISA)proved that the quantities of mmp-2 in experimental group were less than in control group at 1d,3d,7d,1m and 3m after the surgery.The differences were statistically significant(F=4.171,P＜0.05).The quantities of mmp-2 were associated with the loss of the stroma cells and the numbers of spots each(r=0.65,P＜0.05:r=0.58,P＜0.05).The quantities of mmp-9 in experimental group were less than in control group at 1d,3d,7d,1m and 3m after the surgery.The differences were statistically significant(F=5.508,P＜0.05).The quantities of mmp-9 were associated with the loss of the stroma cells and the numbers of spots each(r=0.42,P＜0.05;r=0.31,P＜0.05).3,Zymography for the activity of gelatinase showed that the activity of mmp-2 in experimental group was less than in control group at 1d,3d,7d,1m and 3m after the surgery.The differences were statistically significant(F=4.012,P＜0.05).The activity of mmp-2 were associated with the loss of the stroma cells and the numbers of spots each(r=0.58,P＜0.05: r=0.52,P＜0.05).The activity of mmp-9 in experimental group was less than in control group at 1d,3d,7d,1m and 3m after the surgery.The differences were statistically significant(F=3.822,P＜0.05).The activity of mmp-9 was associated with the lost of the stroma cells and the numbers of spots each(r=0.39,P＜0.05;r=0.28,P＜0.05).4,The result of indirect immuno flurescent for the location of gelatinase show that mmp-2 expressed at the epithelium layer and stroma layer even after LASIK 3m.Maybe mmp-2 has the effect of digesting the basal membrane and normal collagen causing delay of corneal healing.Conclusion:1,Gelatinase A and B were positive association with the lost of stroma cell and the number of spots.GelatinaseA and B may delay the corneal wound healing procession after LASIK.2,The matrix metalloproteinase inhibitor GM6001 could inhibite the expression and activity of gelatinase.May accelerate the corneal wound healing procession after LASIK.