Effect Of Breviscapine On The Epithelial-mesenchymal Transition Induced By TGF-β₂in Human Len Epithelial Cells

BackgroundModern extracapsular cataract extraction or phacoemulsification and posterior chamber intraocular lens implantation is the preferred surgical approach in the world. The residual lens epithelial cells stimulated by various cytokines proliferate, migrate, transdifferentiate in the posterior capsule,which leads to posterior capsular opacification. Posterior capsular opacification is the most common long-term complication after cataract surgery.In recent years, the study of epithelial-mesenchymal transition becomes a hot spot in the eye disease. Epithelial-mesenchymal transition is a major contributor to the pathogenesis of posterior capsular opacification. Studies have shown that Breviscapine has extensive role of anti-fibrosis and suppression epithelial-mesenchymal transition.ObjectivePresent study is to observe the effect of breviscapine on the expression of alpha-smooth muscle actin(a-SMA)and fibronectin(FN) induced by TGF-P2in human len epithelial cells. MethodsAfter HLE-B3was cultured with the different concentration Breviscapine for24、48、72hours. The cell proliferation was detected by CCK-8assay.HLE-B3was cultured in vitro in DMEM containing10%fetal bovine serum and then was divided into4groups. The free-serum culture was used as normal control group.Free-serum culture containing10μg/L TGF-β2was utilized as treatment group.10μg/ml breviscapine was used as Bre-treatment group.10μg/L TGF-β2plused lOug/ml breviscapine was utilized as combination treatment group. After HLE-B3was cultured for72hours, fluorescence quantitative PCR and Western blot were used to detect the mRNA and protein expression of a-SMA and FN.ResultsAfter HLE-B3was cultured with Breviscapine for72hours, the half maximal inhibitory concentration (IC50) of HLE-B3is22μg/ml on CCK-8assay. In this study, the drug concentration lOμg/ml (≈1/2IC50), reaction time was72h to do a follow-up experiment. a-SMA and FN were positively expressed in cultured HLE-B3.The expression levels of a-SMA and FN in HLE-B3were remarkably increased in the group with10μg/L TGF-β2compared with normal control(P=0.000.0.000). The expression levels of a-SMA and FN were obviously declined in combination treatment group in comparison with TGF-β2treatment group (P=0.001、0.001、0.001、0.01). Bre-treatment group was not statistically significant in comparison with normal control(P=0.55、0.292、0.55、0.360).Conclusion1. HLE-B3cultured in vitro can express a certain amount of a-SMA and FN.2. TGF-β2can induce the EMT of lens epithelial cells.3. The certain Breviscapine can inhibit epithelial-mesenchymal transition induced by TGF-β2in human LECs.