Verteporfin without light stimulation inhibits YAP activation in trabecular meshwork cells: Implications for glaucoma treatment
Verteporfin without light stimulation inhibits YAP activation in trabecular meshwork cells: Implications for glaucoma treatment
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DOI：
10.1016/j.bbrc.2015.09.012
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发表时间：
2015-10-16
影响因子：
3.1
通讯作者：
Panjwani, Noorjahan
中科院分区：
文献类型：
--
作者：
Chen, Wei-Sheng;Cao, Zhiyi;Panjwani, Noorjahan
维替普芬是一种光敏剂，用于光动力疗法治疗老年性黄斑变性。在青光眼小鼠模型中，没有光刺激的维替普芬已经被证明可以降低眼压，但其机制尚不清楚。最近的研究表明，维替泊芬在没有光刺激的情况下抑制癌细胞中的YAP。此外，YAP已成为青光眼发病机制中的重要分子。我们推测，在小梁网(TM)中，维替普芬使YAP失活可能与体内观察到的MP减少有关。由于TM组织的收缩能力与眼压有关，因此采用胶原凝胶收缩实验来评价维替普芬对TM细胞收缩能力的影响。将人TM细胞包埋于胶原凝胶中，用维替泊芬处理48h，用ImageJ软件对胶原胶的大小进行定量。为探讨维替泊芬对人TM细胞YAP表达的影响，将维替泊芬作用于人TM细胞24 h，用Western blotting方法检测YAP的表达。为确定维替普芬的细胞毒作用，将维替普芬作用于人TM细胞24小时，然后用WST-1评估细胞活力。我们在这里证明了维替泊芬(I)以剂量依赖的方式取消TM细胞介导的胶原凝胶收缩，(Ii)以剂量依赖的方式抑制YAP和CTGE(结缔组织生长因子，YAP的直接靶基因)的表达，以及(Iii)在2mM以下没有明显的细胞毒性。(C)2015 Elsevier Inc.保留所有权利。
Verteporfin, a photosensitizer, is used in photodynamic therapy to treat age-related macular degeneration. In a glaucoma mouse model, Verteporfin without light stimulation has been shown to reduce intraocular pressure (IOP) but the mechanism is unknown. Recent studies have shown that Verteporfin inhibits YAP without light stimulation in cancer cells. Additionally, YAP has emerged as an important molecule in the pathogenesis of glaucoma. We hypothesize that YAP inactivation by Verteporfin in trabecular meshwork (TM) may be related to the reduced MP observed in vivo. As contractility of TM tissues is associated with IOP, collagen gel contraction assay was used to assess the effect of Verteporfin on contractility of TM cells. Human TM cells were embedded in collagen gel and treated with Verteporfin for 48 h. Areas of collagen gel sizes were quantified by ImageJ. To assess the effect of Verteporfin on the expression of YAP, human TM cells were treated with Verteporfin for 24 h and the expression of YAP was determined by Western blotting. To determine the cytotoxic effect of Verteporfin, human TM cells were treated with Verteporfin for 24 h, and then the cell viability was assessed by WST-1. We demonstrated here that Verteporfin (i) abolishes TM cell-mediated collagen gel contraction in a dose-dependent manner, (ii) attenuates expression of YAP and CTGE (connective tissue growth factor, a direct YAP target gene) in a dose-dependent manner, and (iii) has no significant cytotoxicity below 2 mu M. Taken together, Verteporfin may facilitate aqueous humor outflow through the conventional outflow system and reduce IOP by inactivating YAP. (C) 2015 Elsevier Inc. All rights reserved.