Abstract

Purpose

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.

Methods

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.

Results

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.

Conclusion

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.

中文翻译：

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Eupatilin 减弱 TGF-β2 诱导的视网膜色素上皮细胞增殖和上皮间质转化

摘要

目的

增殖性玻璃体视网膜病变 (PVR) 的主要特征是视网膜色素上皮细胞 (RPE) 的迁移、粘附和上皮间质转化 (EMT)。Eupatilin 是一种天然存在的黄酮，具有抑制细胞增殖和 EMT 的潜力。然而，其对转化生长因子-2 (TGF-β2) 诱导的 PVR 模型的疗效尚不清楚。在这项研究中，研究了 eupatilin 在治疗 RPE 时对增殖和 EMT 的潜在影响。

方法

血清饥饿的人 RPE 细胞 (ARPE-19) 用 10 ng/ml TGF-β2 单独处理或与 25 μM eupatilin 共同处理 48 小时。定量实时 PCR 和蛋白质印迹分析分别用于评估 mRNA 和蛋白质表达水平的目标。通过基于图像的细胞术评估细胞凋亡和细胞周期进程。通过伤口愈合试验评估治疗对细胞迁移的影响。

结果

Eupatilin 通过调节细胞周期和诱导细胞凋亡来抑制 TGF-β2 诱导的 RPE 细胞增殖。TGF-β2 上调间充质标志物纤连蛋白和波形蛋白的 mRNA 表达被处理显着下调，而上皮标志物 E-钙粘蛋白和 occludin 的表达被上调。该疗法通过下调 TGF-β2 诱导的转录因子 Twist、Snail 和 ZEB1 的表达，显着抑制 TGF-β2 促进细胞迁移。此外，eupatilin 显着抑制 MMP-1、-7 和 -9 的表达，并抑制 NF-κB 信号传导。

结论

这些结果表明eupatilin可以抑制RPE细胞的增殖和向成纤维细胞样细胞的转化；因此，该药剂在治疗 PVR 中可能具有潜在的治疗价值。