A Chimeric Cfh Transgene Leads to Increased Retinal Oxidative Stress, Inflammation, and Accumulation of Activated Subretinal Microglia in Mice
A Chimeric Cfh Transgene Leads to Increased Retinal Oxidative Stress, Inflammation, and Accumulation of Activated Subretinal Microglia in Mice
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DOI：
10.1167/iovs.14-16089
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发表时间：
2015-06-01
影响因子：
4.4
通讯作者：
Ufret-Vincenty, Rafael L.
中科院分区：
文献类型：
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作者：
Aredo, Bogale;Li, Tao;Ufret-Vincenty, Rafael L.
目的。影响短共有重复序列 (SCR) 6 至 8 的补体因子 H (Cfh) 变体会增加年龄相关性黄斑变性的风险。我们的目的是使用嵌合 Cfh 转基因小鼠 (chCfhTg) 探讨表达 Cfh 变体对视网膜和 RPE 对氧化应激和炎症的体内敏感性的影响。 chCfhTg 和年龄匹配的 C57BL/6J (B6) 小鼠因正常衰老或暴露于口服氢醌 (0.8% HQ) 和增强光照的组合而遭受氧化应激。收集眼睛用于 RPE-脉络膜平片和视网膜切片的免疫组织化学、ELISA、电子显微镜和 RPE/小胶质细胞基因表达分析。 结果。与 B6 小鼠相比，老化至 2 岁的小鼠导致 chCfhTg 小鼠视网膜中基底层沉积物、视网膜下小胶质细胞/巨噬细胞 (MG/MU) 染色的 CD16 和丙二醛 (MDA) 以及 MDA 修饰蛋白的积累增加。与对照组相比，维持 HQ 饮食和增加光照的 chCfhTg 小鼠表现出更多的基底层沉积物沉积、更多提示 MG/MU 的眼底斑点积累以及 RPE 下空间中 C3d 沉积的增加。此外，chCfhTg 小鼠表现出 RPE/MG/MU 分离 RNA 中 NLRP3、IP-10、CD68 和 TREM-2 的上调。结论。在 SCR 6 至 8 中引入变体的 Cfh 转基因的表达足以导致转基因小鼠模型中视网膜/RPE 对氧化应激的敏感性增加、促炎 MG/MU 表型以及促炎 RPE/MG/MU 基因表达谱。我们的数据表明，Cfh 与 MDA 修饰蛋白相互作用的改变可能与解释 Cfh 变体的作用有关。
PURPOSE. Variants of complement factor H (Cfh) affecting short consensus repeats (SCRs) 6 to 8 increase the risk of age-related macular degeneration. Our aim was to explore the effect of expressing a Cfh variant on the in vivo susceptibility of the retina and RPE to oxidative stress and inflammation, using chimeric Cfh transgenic mice (chCfhTg).METHODS. The chCfhTg and age-matched C57BL/6J (B6) mice were subjected to oxidative stress by either normal aging, or by exposure to a combination of oral hydroquinone (0.8% HQ) and increased light. Eyes were collected for immunohistochemistry of RPE-choroid flat mounts and of retinal sections, ELISA, electron microscopy, and RPE/microglia gene expression analysis.RESULTS. Aging mice to 2 years led to an increased accumulation of basal laminar deposits, subretinal microglia/macrophages (MG/MU) staining for CD16 and for malondialdehyde (MDA), and MDA-modified proteins in the retina in chCfhTg compared to B6 mice. The chCfhTg mice maintained on HQ diet and increased light showed greater deposition of basal laminar deposits, more accumulation of fundus spots suggestive of MG/MU, and increased deposition of C3d in the sub-RPE space, compared to controls. In addition, chCfhTg mice demonstrated upregulation of NLRP3, IP-10, CD68, and TREM-2 in the RNA isolates from RPE/MG/MU.CONCLUSIONS. Expression of a Cfh transgene introducing a variant in SCRs 6 to 8 was sufficient to lead to increased retinal/RPE susceptibility to oxidative stress, a proinflammatory MG/MU phenotype, and a proinflammatory RPE/MG/MU gene expression profile in a transgenic mouse model. Our data suggest that altered interactions of Cfh with MDA-modified proteins may be relevant in explaining the effects of the Cfh variant.