氟轻松通过糖皮质激素受体抑制人视网膜色素上皮（ARPE - 19）细胞中的血管内皮生长因子（VEGF）表达以及鸡胚绒毛尿囊膜（CAM）中肿瘤坏死因子-α（TNF-α）诱导的血管生成。

Fluocinolone inhibits VEGF expression via glucocorticoid receptor in human retinal pigment epithelial (ARPE-19) cells and TNF-alpha-induced angiogenesis in chick chorioallantoic membrane (CAM).

作者信息

Ayalasomayajula Surya P, Ashton Paul, Kompella Uday B

机构信息

University of Nebraska Medical Center, Omaha, Nebraska, USA.

出版信息

J Ocul Pharmacol Ther. 2009 Apr;25(2):97-103. doi: 10.1089/jop.2008.0090.

DOI:10.1089/jop.2008.0090

PMID:19284324

Abstract

PURPOSE

The purpose of this study was to determine whether fluocinolone inhibits vascular endothelial growth factor (VEGF) expression in a retinal pigment epithelial cell line (ARPE-19) and TNF-alpha-induced angiogenesis in chick chorioallantoic membrane (CAM) assay.

METHODS

The dose-dependent effect of fluocinolone (0.0001-1 microM) on VEGF secretion, VEGF mRNA expression, and cytotoxicity was determined in confluent monolayers of ARPE-19 cells using ELISA, RT-PCR, and MTT assay, respectively. The effect of a glucocorticoid receptor antagonist (RU486) on fluocinolone-mediated VEGF expression was determined. The effect of fluocinolone in inhibiting TNF-alpha-induced angiogenesis was determined using chick chorioallantoic membrane (CAM) assay. The dose-dependent effect of fluocinolone (0.0001-1 microM) in inhibiting 1% serum-stimulated ARPE-19 cell proliferation was determined using BrdU labeling assay.

RESULTS

At concentrations devoid of cytotoxicity, fluocinolone inhibited VEGF secretion as well as mRNA expression in ARPE-19 cells. RU486 (1 microM) treatment prevented inhibition of VEGF secretion and VEGF mRNA expression by fluocinolone (0.1 microM). Fluocinolone (50 ng/egg) inhibited angiogenesis induced by TNF-alpha. The ARPE-19 cell proliferation was inhibited by fluocinolone in a dose-dependent manner.

CONCLUSIONS

Fluocinolone inhibited VEGF expression in ARPE-19 cells via its glucocorticoid receptor activity. In addition, fluocinolone inhibited proliferation of ARPE-19 cells and TNF-alpha-induced angiogenesis in chorioallantoic membranes.

摘要

目的

本研究旨在确定氟轻松是否能抑制视网膜色素上皮细胞系（ARPE - 19）中血管内皮生长因子（VEGF）的表达，以及在鸡胚绒毛尿囊膜（CAM）试验中抑制肿瘤坏死因子-α（TNF-α）诱导的血管生成。

方法

分别采用酶联免疫吸附测定（ELISA）、逆转录-聚合酶链反应（RT-PCR）和噻唑蓝（MTT）比色法，测定氟轻松（0.0001 - 1微摩尔/升）对汇合的ARPE - 19细胞单层中VEGF分泌、VEGF信使核糖核酸（mRNA）表达及细胞毒性的剂量依赖性效应。测定糖皮质激素受体拮抗剂（RU486）对氟轻松介导的VEGF表达的影响。采用鸡胚绒毛尿囊膜（CAM）试验，测定氟轻松对TNF-α诱导的血管生成的抑制作用。采用5-溴脱氧尿嘧啶核苷（BrdU）标记法，测定氟轻松（0.0001 - 1微摩尔/升）抑制1%血清刺激的ARPE - 19细胞增殖的剂量依赖性效应。

结果

在无细胞毒性的浓度下，氟轻松抑制ARPE - 19细胞中的VEGF分泌及mRNA表达。RU486（1微摩尔/升）处理可防止氟轻松（0.1微摩尔/升）对VEGF分泌和VEGF mRNA表达的抑制。氟轻松（50纳克/枚鸡蛋）抑制TNF-α诱导的血管生成。氟轻松以剂量依赖性方式抑制ARPE - 19细胞增殖。