Eupatilin attenuates TGF-β2-induced proliferation and epithelial-mesenchymal transition of retinal pigment epithelial cells

• Published in: Cutaneous and ocular toxicology Vol. 40; no. 2; p. 103
• Main Authors: Cinar, Ayca Kupeli, Ozal, S Altan, Serttas, Riza, Erdogan, Suat
• Format: Journal Article
• Language: English
• Published: England 01.06.2021
• Subjects: Antigens, CD - genetics; Cadherins - genetics; Cell Line; Cell Physiological Phenomena - drug effects; Epithelial Cells - drug effects; Epithelial Cells - metabolism; Epithelial-Mesenchymal Transition - drug effects; Fibronectins - genetics; Flavonoids - pharmacology; Humans; Matrix Metalloproteinases - genetics; Nuclear Proteins - genetics; Occludin - genetics; Retinal Pigment Epithelium - cytology; Snail Family Transcription Factors - genetics; Transforming Growth Factor beta2; Twist-Related Protein 1 - genetics; Vimentin - genetics; Vimentin - metabolism; Vitreoretinopathy, Proliferative - drug therapy; Vitreoretinopathy, Proliferative - genetics; Vitreoretinopathy, Proliferative - metabolism; Zinc Finger E-box-Binding Homeobox 1 - genetics; epithelial to mesenchymal transition; TGF-β2; Eupatilin; PVR; retina pigment epithelial cell
• ISSN: 1556-9535
• DOI: 10.1080/15569527.2021.1902343

The main characteristic of proliferative vitreoretinopathy (PVR) is migration, adhesion, and epithelial-mesenchymal transition (EMT) of retinal pigment epithelial cells (RPE). Eupatilin is a naturally occurring flavone that has the potential to inhibit cell proliferation and EMT. However, its efficacy on the PVR model induced by transforming growth factor-2 (TGF-β2) is unknown. In this study, the potential effect of eupatilin on proliferation and EMT in the treatment of RPE was investigated. Serum starved human RPE cells (ARPE-19) were treated with 10 ng/ml TGF-β2 alone or co-treated with 25 μM eupatilin for 48 h. Quantitative real-time PCR and Western blot analysis were used to assess targets at the mRNA and protein expression level, respectively. Apoptosis and cell cycle progression was assessed by image-based cytometry. The effect of treatment on cell migration was evaluated by wound healing assay. Eupatilin inhibited TGF-β2-induced RPE cell proliferation via regulating the cell cycle and inducing apoptosis. TGF-β2 upregulated mRNA expression of mesenchymal markers fibronectin and vimentin was significantly downregulated by the treatment, while the epithelial markers E-cadherin and occludin expression was upregulated. The therapy significantly suppressed TGF-β2 encouraged cell migration through downregulating the expression of transcription factors Twist, Snail, and ZEB1 induced by TGF-β2. Furthermore, eupatilin significantly inhibited the expression of MMP-1, -7, and -9, and suppressed NF-κB signalling. These results suggest that eupatilin could inhibit the proliferation and transformation into fibroblast-like cells of RPE cells; thus the agent may be a potential therapeutic value in treating PVR.