Bromfenac Sodium Inhibits TGF-?2-induced Epithelial-Mesenchymal Transition Via ERK/GSK-3?/Snail Signaling In Human Lens Epithelial Cells

Background and Objective:Phacoemulsification is the only effective method to treat cataract.Posterior capsule opacification(PCO)is the most common long-term complication of modern cataract surgery.The epithelial-mesenchymal transition(EMT)and the processes of proliferation,migration of residual lens epithelial cells(LECs)stimulatedby some growth factors and cytokines is the key pathological mechanism involved in the development PCO.Evidence has shown that TGF-? increasesin postoperative aqueous humor,which is considered to be the main factor of EMT and pathological fibrosis of lens epithelial cells.Studies have shown that TGF-?2 can stimulate the LECs to undergo EMT via ERK pathway.This study was designed to explore the regulation of the process of EMT of LECs induced by TGF-?,and to investigate the potentialmechanism of bromfenac sodium in inhibiting EMT of LECs.Methods:The HLEC-B3 cells were divided into two group according to the absence or presence of TGF-?2(10 ng/ml).fibronectin(FN),matrix metalloproteinase 2(MMP2)and Snail were investigated at mRNA and protein expressionlevels by Real-time fluorescencequantitative PCR and Western blotting assay after treated with TGF-?2 respectively.The migration ability of LECs was measured by cell scratches test,and the morphological changes were observed under optical microscope after stimulated by TGF-?2 for 48h.Cells were pretreated with different concentrations of bromfenac sodium 24 hours prior to TGF-?2 stimulation,and the changes of EMT-related gene expression and the change of migration ability of LECs were detected by Real-time fluorescencequantitative PCR,Western blotting assay,immunofluorescence assay and cell scratches test.To explore the potential mechanism of bromfenac sodium in inhibiting EMT of LECs,Cells were pretreated with bromfenac sodium 24 hours prior to TGF-?2 stimulation,and theproteinexpression levels of p-Smad,p-EKR and p-GSK-3?were investigated by Western blotting assay after treated withTGF-?2.To verify the role of ERK/GSK-3? signaling in EMT of HLEC-B3,the cells were pretreated with MEK specific inhibitor U0126 and GSK-3specific inhibitorCHIR-99021,and detect the expression levels of FN,MMP2 and Snail.To further study the mechanism of ERK/GSK-3? signaling on the regulation of the expression levels of FN and MMP2,the cells were treated with bromfenac sodiumafter transfected with Snail siRNA,and the mRNA and protein expression levels of FN and MMP2were investigated by Real-time fluorescencequantitative PCR and Western blotting assayin order to verify the relationship betweenthe relationship between the inhibitory effect of bromfenac sodium and Snail.Results:The expression levels of FN,MMP2 and Snail were enhanced by the stimulation of TGF-?2 in HLEC-B3.Pretreatment with bromfenac sodium can effectively inhibit TGF-?2-induced cellmigration and overexpression of FN,MMP2 and Snail.TGF-?2 regulates the expressions of FN,MMP2 and Snail via ERK/GSK-3? signaling.Furthermore,bromfenac sodium can inhibit the activation of ERK/GSK-3? signaling induced by TGF-p2in HLEC-B3.Transfection of Snail siRNA inhibits the high expression of FN and MMP2 induced by TGF-?2.Snail siRNA knockdown can effectively limit the inhibitory effect of bromfenac sodium on TGF-?2-induced high expression of FN and MMP2.Conclusion:TGF-?2 can promote cell migration and EMT ofLECs via ERX/GSK-3?/Snail signaling;Bromfenac sodium inhibits TGF-?2-induced cell migration and EMT ofLECs via ERK/GSK-3?/Snail signaling.