Department of Ophthalmology, School of Medicine, Trakya University, Edirne, Turkey.
Department of Medical Biology, School of Medicine, Trakya University, Edirne, Turkey.
Cutan Ocul Toxicol. 2020 Jun;39(2):97-105. doi: 10.1080/15569527.2020.1730882. Epub 2020 Mar 9.
Proliferative vitreoretinopathy (PVR) occurs in approximately 5-10% of patients after retinal detachment surgery. Neferine is a bis-benzylisoquinoline alkaloid found in the green seed embryos () of the lotus flower and has various properties, such as being antithrombotic, antioxidant, neuroprotective, anticancerous, and anti-inflammatory. Although the effects of neferine on the proliferation and migration of cancer cells have been partially shown, their possible role and the mechanism of action on PVR remain unclear. To mimic a PVR model , retinal pigment epithelial (RPE) cells were exposed to epidermal growth factor (EGF) and treated with various concentrations of neferine. Cell viability was determined by MTT test. Cell-cycle phase distribution and cell migration were examined by image-based cytometry and wound healing test, respectively. Messenger RNA (mRNA) and protein expression were determined by RT-qPCR and Western blotting, respectively. Stimulation of the cells with EGF significantly increased the rate of proliferation, whilst treatment with low concentrations of neferine-reduced proliferation to a level equal to that seen in untreated cells. Neferine significantly downregulated EGF-increased cell viability, and survivin mRNA expression was depressed to the basal level. In addition, neferine treatment contributed to cell proliferation loss by upregulating p21 and p27 expression leading to cycle arrest at the G1 phase. The treatment significantly inhibited cell migration by upregulating the expression of epithelial markers, such as E-cadherin and occludin, and decreased MMP2, MMP9, α-SMA, and vimentin. Neferine treatment markedly reduced phosphotidyl inositol 3-kinase (PI3K), AKT, p-p38 mitogen-activated protein kinase (MAPK), and NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) protein expression. It can be considered that neferine may be a potential candidate molecule in the treatment of PVR by inhibiting cell proliferation and the migration of EGF-induced RPE cells through the modulation of various transcriptional activities.
增殖性玻璃体视网膜病变（PVR）在视网膜脱离手术后约 5-10%的患者中发生。莲心碱是从荷叶的绿色种胚中提取的一种双苄基异喹啉生物碱，具有抗血栓形成、抗氧化、神经保护、抗癌和抗炎等多种特性。虽然已经部分显示了莲心碱对癌细胞增殖和迁移的影响，但它们在 PVR 中的可能作用和作用机制尚不清楚。为了模拟 PVR 模型，将视网膜色素上皮（RPE）细胞暴露于表皮生长因子（EGF）并以不同浓度的莲心碱处理。通过 MTT 试验测定细胞活力。通过图像细胞计数法和划痕愈合试验分别检查细胞周期相分布和细胞迁移。通过 RT-qPCR 和 Western blotting 分别测定信使 RNA（mRNA）和蛋白质表达。EGF 刺激细胞显著增加增殖率，而低浓度莲心碱处理将增殖降低至未处理细胞的水平。莲心碱显著下调 EGF 增加的细胞活力，并将 survivin mRNA 表达下调至基础水平。此外，莲心碱处理通过上调 p21 和 p27 表达导致细胞周期停滞在 G1 期，从而导致细胞增殖丧失。该治疗方法通过上调上皮标志物（如 E-钙粘蛋白和封闭蛋白）的表达显著抑制细胞迁移，并降低 MMP2、MMP9、α-SMA 和波形蛋白的表达。莲心碱处理显著降低了磷酯酰肌醇 3-激酶（PI3K）、AKT、p-p38 丝裂原激活蛋白激酶（p38 MAPK）和 NF-κB（核因子 kappa-轻链增强子的激活 B 细胞）蛋白表达。可以认为，莲心碱可能通过调节各种转录活性来抑制 EGF 诱导的 RPE 细胞的增殖和迁移，成为治疗 PVR 的潜在候选分子。
