Suppression of Choroidal Neovascularization by N-Acetyl-Cysteine in Mice
Suppression of Choroidal Neovascularization by N-Acetyl-Cysteine in Mice
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DOI：
10.3109/02713683.2010.500112
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发表时间：
2010-11-01
影响因子：
2
通讯作者：
Tanihara, Hidenobu
中科院分区：
文献类型：
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作者：
Hara, Ryuhei;Inomata, Yasuya;Tanihara, Hidenobu
方法：采用激光光凝法建立C57 BL/6小鼠CNV模型。将小鼠腹膜内注射NAC或单独的媒介物。Westernblotting法检测4-羟基-2-壬烯醛（4-HNE）修饰蛋白和核因子（NF）-κ B（B）的表达。激光损伤后巨噬细胞和中性粒细胞的募集进行了化学分析和髓过氧化物酶（MPO）测定。采用酶联免疫吸附试验（ELISA）测定单核细胞趋化蛋白（MCP）-1、CXCL 1、血管内皮生长因子（VEGF）、VEGF受体（VEGFR）-1和VEGFR-2。CNV的程度进行了评估后7天激光损伤凝集素staining.Results：在NAC治疗的小鼠与激光诱导的injuries. Results，诱导4-HNE-修饰的蛋白3小时后，核提取物中的NF-κ B的激活后6小时显着抑制相比，车辆治疗的小鼠。与溶剂处理的小鼠相比，NAC处理的小鼠中巨噬细胞和中性粒细胞募集受到抑制，MCP-1、CXCL 1、VEGF和VEGFR-1的水平也较低。此外，在NAC处理的小鼠中诱导的CNV的程度显著低于溶剂处理的小鼠（p = 0.027）。结论：我们的结果清楚地表明，NAC抑制氧化应激的指标和由激光损伤诱导的NF-κ B的活化，并且因此抑制巨噬细胞和中性粒细胞浸润以及CNV的发展。这为年龄相关性黄斑变性提出了新的预防和干预治疗策略。
Methods: CNV was induced in C57BL/6 mice by laser photocoagulation of the ocular fundus. Mice were injected intraperitoneally with NAC or vehicle alone. The levels of 4-hydoroxy-2-nonenal (4-HNE)-modified protein and nucleus factor (NF)-kappa B were determined by wester blotting. The recruitment of macrophages and neutrophils after laser injury was analyzed immunohistochemically and in myeloperoxidase (MPO) assays. Enzyme-linked immunosorbent assays (ELISA) were used to measure monocyte chemotactic protein (MCP)-1, CXCL1, vascular endothelial growth factor (VEGF), VEGF receptor (VEGFR)-1, and VEGFR-2. The extent of CNV was evaluated 7 d after laser injury by lectin staining.Results: In NAC-treated mice with laser-induced injuries, the induction of 4-HNE-modified protein after 3 hr and the activation of NF-kappa B in nuclear extracts after 6 hr were markedly suppressed compared to vehicle-treated mice. Macrophage and neutrophil recruitment were inhibited and the levels of MCP-1, CXCL1, VEGF, and VEGFR-1 were also lower in NAC-treated mice compared to vehicle-treated mice. Furthermore, the extent of CNV induced was significantly lower in NAC-treated compared to vehicle-treated mice (p = 0.027).Conclusions: Our results clearly showed that NAC inhibited indicators of oxidative stress and the activation of NF-kappa B induced by laser injury, and, consequently, suppressed macrophage and neutrophil infiltration and the development of CNV. This suggests novel preventative and interventional therapeutic strategies for age-related macular degeneration.