Complement Stimulates Retinal Pigment Epithelial Cells to Undergo Pro-Inflammatory Changes.
Complement Stimulates Retinal Pigment Epithelial Cells to Undergo Pro-Inflammatory Changes.
复制标题
补体刺激视网膜色素上皮细胞发生促炎症变化。
DOI：
10.1159/000439596
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发表时间：
2015
影响因子：
2.1
通讯作者：
Lueck K
中科院分区：
文献类型：
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作者：
Lueck K
背景/目的我们研究了人补体血清(HCS)对视网膜色素上皮(RPE)细胞的影响，以及与早期老年性黄斑变性(AMD)相关的促炎介质的作用。用免疫细胞化学和流式细胞术分析细胞C5b-9。采用ELISA法和RT-PCR法测定IL-6、IL-8和单核细胞趋化蛋白-1(MCP-1)的含量。免疫印迹法检测肿瘤坏死因子-α(TNF-a)、细胞间黏附分子-1(ICAM-1)和血管细胞黏附分子-1(VCAM-1)。结果经κ处理后，C5b-9染色呈浓度依赖性增加，但对细胞存活率无明显影响。酶联免疫吸附试验和RT-PCR检测显示IL-6、IL-8和MCP-1的分泌和表达增加。免疫荧光染色显示核因子κB的核转位。结论补体可促进视网膜色素上皮细胞中核因子κB的活化，从而可能进一步创造促炎环境。所有这些因素加在一起可能会支持AMD的早期发展。
Background/AimsWe examined the effect of human complement sera (HCS) on retinal pigment epithelial (RPE) cells with respect to pro-inflammatory mediators relevant in early age-related macular degeneration (AMD).MethodsRPE cells were treated with complement-containing HCS or with heat-inactivated (HI) HCS or C7-deficient HCS as controls. Cells were analysed for C5b-9 using immunocytochemistry and flow cytometry. Interleukin (IL)-6, IL-8, and monocyte chemoattractant protein-1 (MCP-1) were quantified by ELISA and RT-PCR. Tumour necrosis factor-a (TNF-a), intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1), were analysed by Western blotting. The intracellular distribution of nuclear factor (NF)-κB was investigated by immunofluorescence.ResultsA concentration-dependent increased staining for C5b-9 but no influence on cell viability was observed after HCS treatment. ELISA and RT-PCR analysis revealed elevated secretion and expression of IL-6, IL-8, and MCP-1. Western blot analysis showed a concentration-dependent increase in ICAM-1, VCAM-1, and TNF-a in response to HCS, and immunofluorescence staining revealed nuclear translocation of NF-κB.ConclusionThis study suggests that complement stimulates NF-κB activation in RPE cells that might further create a pro-inflammatory environment. All these factors together may support early AMD development.