Effects of mechanical stress and vitreous samples in retinal pigment epithelial cells.
Effects of mechanical stress and vitreous samples in retinal pigment epithelial cells.
复制标题
DOI：
10.1016/j.bbrc.2016.01.104
复制
发表时间：
2016-02
影响因子：
3.1
通讯作者：
Eri Takahashi;Ayako Fukushima;A. Haga;Yasuya Inomata;Yasuhiro Ito;M. Fukushima;H. Tanihara
中科院分区：
文献类型：
--
作者：
Eri Takahashi;Ayako Fukushima;A. Haga;Yasuya Inomata;Yasuhiro Ito;M. Fukushima;H. Tanihara
在孔源性视网膜脱离（RRD）中，分散的RPE细胞从基底膜进入玻璃体腔，经历上皮间质转化（EMT），并形成眼内纤维膜，以响应玻璃体液。从患有RRD、视网膜前膜（ERM）或黄斑裂孔（MH）的患者收集人玻璃体样品。我们评估了玻璃体对ARPE-19细胞的影响，在悬浮培养中使用聚甲基丙烯酸2-羟乙酯包被的培养皿和三维（3D）Matrigel培养。我们发现，暴露于玻璃体样本不会引起形态学变化或加速单层伤口闭合。一些样品显示Smad 2磷酸化和核因子-κB核转位增加。机械应力引发Smad 2磷酸化水平升高。此外，暴露于玻璃体液增加了机械应力后悬浮培养细胞中p38丝裂原活化蛋白激酶的磷酸化。此外，ARPE-19细胞在3D Matrigel培养中显示出星状侵袭表型，在这项研究中，我们证明了机械应力和玻璃体与3D培养中的EMT相关信号和侵袭表型相关，而不是在单层中。这些结果对玻璃体液在诱导EMT和眼内纤维化中的作用具有重要意义。
In rhegmatogenous retinal detachment (RRD), scattered RPE cells from the basement membrane into the vitreous cavity undergo an epithelial mesenchymal transition (EMT) and form the intraocular fibrous membrane in response to vitreous fluid.We investigated whether exposure to vitreous samples was associated with EMT-associated signals and mesenchymal characters. Human vitreous samples were collected from patients with RRD, epiretinal membrane (ERM), or macular hole (MH). We evaluated the effects of vitreous on ARPE-19 cells in suspension cultures using poly 2-hydroxyethyl methacrylate-coated dishes and three-dimensional (3D) Matrigel cultures. We found that exposure to vitreous samples did not induce morphological changes or accelerate wound closure in monolayers. Several samples showed increased phosphorylation of Smad2 and nuclear translocation of nuclear factor-κB. Mechanical stress triggered an elevation of phosphorylation levels in Smad2. In addition, exposure to vitreous fluid increased the phosphorylation of p38 mitogen-activated protein kinase in cell suspension cultures after mechanical stress. Moreover, ARPE-19 cells showed a stellate invasive phenotype in 3D Matrigel cultures with vitreous samples.In this study, we demonstrated that mechanical stress and vitreous were associated with EMT-associated signals and invasive phenotypes in 3D cultures but not in monolayers. These results have important implications for the role of vitreous humor in the induction of EMT and intraocular fibrosis.