Effects of Secreted Mast Cell Mediators on Retinal Pigment Epithelial Cells: Focus on Mast Cell Tryptase.

Effects of Secreted Mast Cell Mediators on Retinal Pigment Epithelial Cells: Focus on Mast Cell Tryptase.
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DOI:
10.1155/2017/3124753
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发表时间:
2017
影响因子:
4.6
通讯作者:
Ebihara N
Ebihara N
中科院分区:
医学3区
文献类型:
--
作者:
Arai R;Usui-Ouchi A;Ito Y;Mashimo K;Murakami A;Ebihara N

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脉络膜中存在大量肥大细胞,但肥大细胞介质对视网膜色素上皮(RPE)细胞的影响尚不清楚。我们研究了肥大细胞介质在体外对RPE细胞的影响,重点是类胰蛋白酶。通过逆转录聚合酶链反应检测受体的表达。我们还评估了白细胞介素8和血管内皮生长因子(VEGF)的生产后,RPE细胞刺激肥大细胞介质通过使用抗体阵列和酶联免疫吸附试验。此外,我们研究了类胰蛋白酶对RPE细胞迁移和完整性的影响,通过划痕实验和跨上皮电阻。RPE细胞表达蛋白酶激活受体2(PAR 2)、组胺受体1、肿瘤坏死因子-α(TNF-α)受体1和CCR 1、3、4、8和11。类胰蛋白酶、PAR 2激动剂、组胺和TNF-α均能促进RPE细胞产生白细胞介素8,而只有类胰蛋白酶能促进VEGF的产生。类胰蛋白酶还增强磷酸化细胞外信号调节激酶1/2的表达,导致RPE细胞迁移增加。然而,类胰蛋白酶并没有改变上皮的完整性或表达的封闭小带-1和连接粘附分子-A的RPE细胞。肥大细胞介质,特别是类胰蛋白酶,可能影响RPE细胞炎症。
Numerous mast cells are present in the choroid, but the effects of mast cell mediators on retinal pigment epithelial (RPE) cells are not well understood. We investigated the influence of mast cell mediators on RPE cells in vitro, focusing on tryptase. Expression of receptors was examined by the reverse transcription polymerase chain reaction. We also assessed production of interleukin 8 and vascular endothelial growth factor (VEGF) after RPE cells were stimulated with mast cell mediators by using an antibody array and enzyme-linked immunosorbent assay. Furthermore, we investigated the influence of tryptase on RPE cell migration and integrity by the scratch assay and the transepithelial resistance. RPE cells expressed protease-activated receptor 2 (PAR2), histamine receptor 1, tumor necrosis factor-α (TNF-α) receptor 1, and CCR 1, 3, 4, 8, and 11. Tryptase, PAR2 agonists, histamine, and TNF-α all enhanced interleukin 8 production by RPE cells, while only tryptase enhanced VEGF production. Tryptase also enhanced expression of phosphorylated extracellular signal-regulated kinases 1/2, resulting in increased migration of RPE cells. However, tryptase did not alter epithelial integrity or the expression of zonula occludens-1 and junctional adhesion molecule-A by RPE cells. Mast cell mediators, especially tryptase, may influence RPE cell inflammation.
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