上海交通大学学报(医学版) ›› 2019, Vol. 39 ›› Issue (6): 571-.doi: 10.3969/j.issn.1674-8115.2019.06.003

• 论著·基础研究 • 上一篇    下一篇

叶黄素对转化生长因子 -β2诱导的视网膜色素上皮细胞 上皮 -间质转化的影响

吕亚男,顾青,李东丽,宫媛媛   

  1. 上海交通大学附属第一人民医院眼科,上海市眼底病重点实验室,上海市眼视觉及光医学工程研究中心,上海 200080
  • 出版日期:2019-06-28 发布日期:2019-07-26
  • 通讯作者: 宫媛媛,电子信箱:gyydr@126.com。
  • 作者简介:吕亚男(1993—),女,硕士生;电子信箱: lvyanan@sjtu.edu.cn。
  • 基金资助:
    国家重点研发计划(2016YFC0904800);上海市第一人民医院临床研究创新团队建设项目(CTCCR-2018BP04)

Effects of lutein on transforming growth factor-β2 induced epithelial-mesenchymal transition in ARPE-19 cells

Lü Ya-nan, GU Qing, LI Dong-li, GONG Yuan-yuan   

  1. Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai Key Laboratory of Ocular Fundus Disease, Department of Ophthalmology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080, China
  • Online:2019-06-28 Published:2019-07-26
  • Supported by:
    National Key R&D Plan of China, 2016YFC0904800; Clinical Research Innovation Plan of Shanghai General Hospital, CTCCR-2018BP04)。

摘要: 目的 ·建立转化生长因子 -β2(transforming growth factor-β2,TGF-β2)诱导的人视网膜色素上皮细胞(retinal pigment epithelium cell,RPE)上皮 -间质转化(epithelial-mesenchymal transformation,EMT)模型,探讨叶黄素的作用及其机制。方法 ·体外培养 ARPE-19细胞,分为空白组、 TGF-β2组、TGF-β2+叶黄素组、叶黄素组。采用 real-time PCR检测各组细胞 α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、纤连蛋白(fibronectin,FN)、上皮钙黏素(E-cadherin)mRNA的表达。 Western blotting检测各组细胞中 α-SMA、FN、闭合蛋白(occludin)的蛋白表达。采用免疫荧光检测 α-SMA的表达。同时用 Western blotting检测 TGF/ Smad通路下游 Smad3的磷酸化水平。结果 · TGF-β2+叶黄素组 EMT程度有明显减轻。纤维化指标 α-SMA、FN的 mRNA、蛋白表达下降,与 TGF-β2组相比差异有统计学意义(均 P<0.05)。同时,上皮细胞相关指标 E-cadherin mRNA、occludin蛋白的表达上调(均 P<0.05)。免疫荧光结果显示,叶黄素可明显抑制上皮细胞转化为肌成纤维细胞。此外, TGF-β2诱导 ARPE-19细胞 EMT过程中发生 Smad3磷酸化水平升高现象可被叶黄素干预(P0.001)。结论 ·叶黄素通过调控 TGF-β/Smad信号通路抑制 ARPE-19细胞 EMT过程,具有潜在的抑制视网膜下纤维化的作用。

关键词: 视网膜下纤维化, 叶黄素, 转化生长因子 -&, beta, 2, 上皮 -间质转化, 视网膜色素上皮细胞

Abstract: Objective · To establish the transforming growth factor-β2 (TGF-β2) induced epithelial-mesenchymal transition (EMT) model of retinal pigment epithelium cells, and investigate the effect and mechanism of lutein on EMT. Methods · ARPE-19 cells were cultured and divided into 4 groups including control group, TGF-β2 group, TGF-β2+lutein group and lutein group. The mRNA levels of α-smooth muscle actin (α-SMA), fibronectin (FN) and E-cadherin were analyzedreal-time PCR. The protein of α-SMA, FN and occludin were assayedWestern blotting. Immunofluorescence was used to detect the change of α-SMA. Meanwhile, Western blotting was performed to detect the levels of pSmad3 in the TGF/Smad signaling pathway. Results · TGF-β2 induced EMT was inhibitedlutein. Lutein decreased the mRNA and protein levels of the mesenchymal markers α-SMA and FN, and increased the of the epithelial markers E-cadherin and occludin (all P<0.05). Immunofluorescence showed that lutein can inhibit the conversion of epithelial cells into myofibroblasts. Lutein significantly downregulated the high of pSmad3 in TGF-β2 treated ARPE-19 cells (P0.001). Conclusion · Lutein inhibits TGF-β2 induced EMTdownregulating the of pSmad3 in TGF-β/ Smad signaling pathway, indicating it may attenuate subretinal fibrosis.

Key words: subretinal fibrosis, lutein, transforming growth factor-&, beta, 2 (TGF-&, beta, 2), epithelial-mesenchymal transformation (EMT), retinal pigment epithelium cell (RPE)

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