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人类视网膜色素上皮细胞表达长链五聚体蛋白PTX3。

Human retinal pigment epithelial cells express the long pentraxin PTX3.

作者信息

Woo Je Moon, Kwon Min-Young, Shin Da-Yong, Kang Young-Ho, Hwang Narae, Chung Su Wol

机构信息

Department of Opthalmology, Ulsan University Hospital, Ulsan, South Korea.

出版信息

Mol Vis. 2013;19:303-10. Epub 2013 Feb 6.

Abstract

PURPOSE

To determine whether the long pentraxin 3 (PTX3) is expressed in human retinal pigment epithelial cells and is induced by inflammatory cytokines, interleukin-1 beta (IL-1β), tumor necrosis factor-alpha (TNF-α), and interferon-gamma (IFN-γ), expression of PTX3 was investigated in the human retinal pigment epithelial cell line, ARPE-19 cells.

METHODS

In ARPE-19 cells, we first analyzed PTX3 production in the presence or absence of inflammatory cytokines, IL-1β, TNF-α, and IFN-γ, dose- and time-dependently using enzyme-linked immunosorbent assay. Protein and mRNA expression of PTX3 was measured with western blotting analysis and real-time reverse transcription-polymerase chain reaction. Specific inhibitors were used to determine the signaling pathways of inflammatory cytokine-induced PTX3 expression.

RESULTS

In this study, production of PTX3 was induced by IL-1β and TNF-α dose- and time-dependently, but not by IFN-γ in ARPE-19 cells. Protein and mRNA expression of PTX3 was significantly upregulated in the presence of IL-1β and TNF-α. Furthermore, pretreatment with extracellular signal-regulated kinase1/2 and nuclear factor kappa-light-chain-enhancer of activated B cells specific inhibitor abolished IL-1β and TNF-α-induced PTX3 production, but the other inhibitors had no effect.

CONCLUSIONS

These results suggested that human retinal pigment epithelial cells may be a major source of PTX3 production in the presence of proinflammatory cytokines, IL-1β and TNF-α, and could be an important mediator for host defense and inflammatory response in the retina. The importance of the mitogen-activated protein kinase/extracellular signal-regulated kinase1/2 and nuclear factor kappa-light-chain-enhancer of activated B cells pathways for regulated PTX3 expression may be a potential target for PTX3 regulation in the retina.

摘要

目的

为了确定长五聚体蛋白3(PTX3)是否在人视网膜色素上皮细胞中表达,以及是否受炎性细胞因子白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)和干扰素-γ(IFN-γ)诱导,我们在人视网膜色素上皮细胞系ARPE-19细胞中研究了PTX3的表达情况。

方法

在ARPE-19细胞中,我们首先使用酶联免疫吸附测定法,剂量和时间依赖性地分析了在存在或不存在炎性细胞因子IL-1β、TNF-α和IFN-γ的情况下PTX3的产生。用蛋白质印迹分析和实时逆转录-聚合酶链反应测量PTX3的蛋白质和mRNA表达。使用特异性抑制剂来确定炎性细胞因子诱导的PTX3表达的信号通路。

结果

在本研究中,ARPE-19细胞中PTX3的产生受IL-1β和TNF-α剂量和时间依赖性诱导,但不受IFN-γ诱导。在存在IL-1β和TNF-α的情况下,PTX3的蛋白质和mRNA表达显著上调。此外,用细胞外信号调节激酶1/2和活化B细胞核因子κ轻链增强子特异性抑制剂预处理可消除IL-1β和TNF-α诱导的PTX3产生,但其他抑制剂无效。

结论

这些结果表明,在促炎细胞因子IL-1β和TNF-α存在的情况下,人视网膜色素上皮细胞可能是PTX3产生的主要来源,并且可能是视网膜中宿主防御和炎症反应的重要介质。丝裂原活化蛋白激酶/细胞外信号调节激酶1/2和活化B细胞核因子κ轻链增强子通路对PTX3表达的调节作用的重要性,可能是视网膜中PTX3调节的潜在靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6b91/3566900/e2698ed39c3e/mv-v19-303-f1.jpg

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