| Purpose:Glaucoma is the first irreversible blinding eye disease with progressive damage to optic nerve and visual field.And the filtering surgery,up till now,is the main surgical treatment for the medium and advanced patients.However,the excessive healing and fibrosis of the filtering passage can result in the failure,which has forced many patients to undergo the reoperation.Human conjunctival fibroblasts(Hcon F)are of great significance in the formation of the scar tissue in filtering passage,that can be stimulated to proliferate,transform into myofibroblasts and synthesize more ECM by TGF-?.Even if the traditional anti-metabolic drugs paly an important role in inhibiting the scarring,filtering bleb leaking,corneal endothelium decompensation,delayed endophthalmitis and other severe complications can be caused easily.What's more,the stop production of Mitomycin C in the early period in China has also hindered the anti-scarring treatment(Production has been resumed now).Therefore,finding an effective,safe and convenient anti-scarring drug has become a long-term goal to improve the success rate of the surgery.Bromfenac sodium is a non-steroidal anti-inflammatory drug used for the treatment of inflammation in the eyes.However,studies have found that bromfenac sodium can inhibit TNF-?-induced proliferation and migration of human lens epithelial cells by inhibiting COX-2,in addition to anti-inflammatory function.Therefore,bromfenac sodium was applied to human conjunctival fibroblasts to evaluate its role in fibrotic processes of human conjunctival fibroblasts in this study.Methods:Human conjunctival fibroblasts were induced by TGF-?1 to establish the model after glaucoma filtering surgery in vitro.Cell proliferation was determined by CCK-8assay.Western blotting assays and cell immunofluorescence staining were performed to detect the expression of ?-SMA,fibronectin,collagen I and COX-2,respectively.Results:CCK-8 assay showed that bromfenac sodium at different concentrations(40,100,200,400 ?g/ml)inhibited the proliferation of Hcon F in a concentration-dependent manner compared with the blank control group(P<0.01).The differences between the group of 40 ?g/ml and the groups with different dose(100,200,400?g/ml)were statistically significant(P<0.01),but there were no statistical differences between the groups of 100 ?g/ml and 200 ?g/ml,200 ?g/ml and400 ?g/ml.So 100?g/ml was selected as the final concentration of bromfenac sodium.Then bromfenac sodium was applied to TGF-?1-induced Hcon F.Compared with the group of TGF-?1,there was fewer cell counts in the group of TGF-?1+BS(P<0.01).The results of western blotting and immunofluorescence staining showed that the expression of ?-SMA protein decreased in the group of TGF-?1+BS,compared with the group of TGF-?1(P<0.01).Similarly,the expression of fibronectin and collagen I in group of TGF-?1+BS were lower than that in the group of TGF-?1(fibronectin P<0.05,collagen-I P <0.001).And there were no statistical differences between the groups of control and TGF-?1 in the COX-2 protein expression.However,the expression of COX-2 protein in the group of TGF-?1+BS was significantly increased compared with the group of TGF-?1(P < 0.01).Conclusions:Bromfenac sodium can inhibit the fibrotic processes,including proliferation,transformation and extracellular matrix formation,of human conjunctival fibroblasts induced by TGF-?1,which may be related to the increased expression of COX-2. |
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