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白细胞介素-17对人视网膜血管内皮细胞体外毛细血管管形成的影响。

Effects of interleukin-17 on human retinal vascular endothelial cell capillary tube formation in vitro.

作者信息

Liu Gaoqin, Wu Hongya, Chen Lei, Xu Jing, Wang Mengjiao, Li Dan, Lu Peirong

机构信息

Department of Ophthalmology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215006, P.R. China.

Jiangsu Clinical Immunology Institute, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215006, P.R. China.

出版信息

Mol Med Rep. 2017 Jul;16(1):865-872. doi: 10.3892/mmr.2017.6623. Epub 2017 May 25.

Abstract

The present study aimed to investigate the effect of and mechanism underlying interleukin (IL)‑17 on human retinal vascular endothelial cell (HREC) capillary tube formation in vitro. The expression of IL‑17 receptor (IL‑17R) in human HRECs was quantified using reverse transcriptase‑polymerase chain reaction (RT‑PCR) and western blot analyses. The roles of IL‑17 in HREC migration and capillary tube formation were detected using a wound scratching assay and three‑dimensional Matrigel assay, respectively, in vitro. HREC proliferation was examined using a cell counting kit‑8 assay with administration of serial doses of IL‑17. The effects of IL‑17 on the expression of vascular endothelial growth factor (VEGF), intercellular cell adhesion molecule (ICAM)‑1, IL‑6 and IL‑8 in HRECs were evaluated using RT‑PCR and western blot analyses. The results revealed that the HRECs expressed IL‑17R, and the number of intact capillary tubes formed by HRECs in the presence of IL‑17 was markedly higher, compared with that in the blank control group. The wound scratching assay showed that the numbers of migrated HRECs stimulated with IL‑17 at concentrations of 100 or 500 ng/ml were significantly higher, compared with the number in the control group. The RT‑PCR and western blot analyses showed that IL‑17 significantly promoted the expression of VEGF, ICAM‑1, IL‑6 and IL‑8 by the HRECs. The proliferation of HRECs in the presence of IL‑17 was also significantly increased. Therefore, IL‑17 increased HREC capillary tube formation through promoting HREC migration, proliferation, and expression levels of VEGF, ICAM‑1, IL‑6 and IL-8.

摘要

本研究旨在探讨白细胞介素(IL)-17对人视网膜血管内皮细胞(HREC)体外形成毛细血管管的作用及其潜在机制。采用逆转录-聚合酶链反应(RT-PCR)和蛋白质免疫印迹分析对人HRECs中IL-17受体(IL-17R)的表达进行定量。分别采用划痕试验和三维基质胶试验在体外检测IL-17在HREC迁移和毛细血管管形成中的作用。使用细胞计数试剂盒-8法并给予系列剂量的IL-17检测HREC增殖情况。采用RT-PCR和蛋白质免疫印迹分析评估IL-17对HRECs中血管内皮生长因子(VEGF)、细胞间黏附分子(ICAM)-1、IL-6和IL-8表达的影响。结果显示,HRECs表达IL-17R,与空白对照组相比,在IL-17存在的情况下HREC形成的完整毛细血管管数量明显更多。划痕试验表明,浓度为100或500 ng/ml的IL-17刺激后迁移的HREC数量显著高于对照组。RT-PCR和蛋白质免疫印迹分析表明,IL-17显著促进HRECs中VEGF、ICAM-1、IL-6和IL-8的表达。在IL-17存在的情况下HREC的增殖也显著增加。因此,IL-17通过促进HREC迁移、增殖以及VEGF、ICAM-1、IL-6和IL-8的表达水平增加HREC毛细血管管的形成。

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