The Ingenious Interactions Between Macrophages and Functionally Plastic Retinal Pigment Epithelium Cells
The Ingenious Interactions Between Macrophages and Functionally Plastic Retinal Pigment Epithelium Cells
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巨噬细胞与功能性可塑性视网膜色素上皮细胞之间的巧妙相互作用
DOI：
10.1167/iovs.16-20604
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Hamuro J.
中科院分区：
文献类型：
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作者：
Yamawaki T;Ito E;Mukai A;Ueno M;Yamada J;Sotozono C;Kinoshita S;Hamuro J.
目的：方法：将贴壁的腹膜细胞或小鼠MP细胞系Raw 264.7与C57 BL/6小鼠原代RPE细胞共培养，观察在LPS或TNF-α刺激或不刺激条件下，MP与RPE细胞的相互作用。然后用Bio-Rad鼠23-panel测定试剂盒（Bio-Rad Laboratories）分析培养上清液（CS）的细胞因子水平。ELISA法检测CS中单核细胞趋化蛋白-1（MCP-1）、IL-6、VEGF和TNF-α的含量。在共培养物中，通过定量实时PCR分析补体相关基因、TNF-α和血管生成相关基因的表达谱。7细胞与RPE细胞共培养，并与MPs和RPE细胞单独培养的细胞进行比较。在没有直接细胞接触的情况下证实了协同效应，并且在LPS或TNF-α存在下更加突出。RPE细胞抑制MP产生TNF-α。RPE细胞与RAW 264的共培养。7细胞C3、CFB和VEGF基因表达增加，而补体调节因子CFH、CD 59、clusterin、TNF-α和抗血管生成色素上皮衍生因子（PEDF.Conclusions）基因表达减少。
Purpose: The purpose of this study was to clarify the interactions between macrophages (MPs) and RPE cells in coculture systems to investigate the functional plasticity of RPE cells.Methods: Adherent peritoneal cells or murine MP cell line Raw 264.7 was cocultured with primary RPE cells taken from C57BL/6 mice, with or without lipopolysaccharide (LPS) or TNF-α stimulation. The cytokine levels of the culture supernatants (CSs) were then analyzed with the Bio-Plex murine 23-Plex Panel Assay Kit (Bio-Rad Laboratories). Monocyte chemoattractant protein-1 (MCP-1), IL-6, VEGF, and TNF-α in CS were further quantified by ELISA. The expression profiles, in cocultures, of complement-associated genes, TNF-α, and angiogenesis-associated genes were analyzed by quantitative real-time PCR.Results: The production of MCP-1, IL-6, and VEGF was synergistically elevated when primary MPs or RAW264. 7 cells and RPE cells were cocultured compared with those derived from sole cultures of MPs and RPE cells. The synergistic effect was confirmed without direct cell contact and was more prominent in the presence of LPS or TNF-α. TNF-α production by MPs was suppressed by RPE cells. Coculture of RPE cells with RAW264. 7 cells increased the gene expression of C3, CFB, and VEGF genes, whereas it reduced those of complement regulatory factors CFH, CD59, clusterin, and TNF-α and antiangiogenic pigment epithelium-derived factor (PEDF).Conclusions: Our findings indicate the presence of ingenious interactions between MPs and RPE cells that forces the inflammation and complement activation in the vicinity of RPE cells under pathologic conditions.