Inomata Yasuya, Tanihara Hidenobu, Tanito Masaki, Okuyama Hiroaki, Hoshino Yuma, Kinumi Tomoya, Kawaji Takahiro, Kondo Norihiko, Yodoi Junji, Nakamura Hajime
Department of Ophthalmology and Visual Science, Kumamoto University Graduate School of Medical Sciences, Kumamoto, Japan.
Invest Ophthalmol Vis Sci. 2008 Nov;49(11):5118-25. doi: 10.1167/iovs.07-1659. Epub 2008 May 30.
To examine the role of thioredoxin-1 (TRX-1), an endogenous protein with a variety of redox-related roles, in the formation of choroidal neovascularization (CNV).
CNV was induced by laser photocoagulation of the ocular fundus in wild-type and transgenic mice overexpressing human TRX-1 (TRX-1 Tg). Mice were injected intraperitoneally with TRX-1, mutant TRX, or vehicle. The incidence of CNV was evaluated by lectin staining. Leukocyte recruitment and C3b deposition after laser injury were determined by immunohistochemistry and Western blotting. Moreover, TRX-1-associated proteins from human plasma were isolated by two-dimensional gel electrophoresis with the use of a column coupled with a mutant TRX-1 and were identified by mass spectrometry and proteomics analysis. Complement activation was determined by a fluid-phase
The incidence of laser-induced CNV was reduced in TRX-1 Tg mice (56.1%) and in C57B/6 mice treated with TRX-1 (46.7%) but not in mutant TRX-1 (79.2%) compared with wild-type mice (85.7%). Furthermore, leukocyte recruitment was prevented in TRX-1-treated mice; C3b deposition was decreased in these and TRX-1 Tg mice. In human plasma, five proteins associated with TRX-1 were identified as apolipoprotein A-I, the CD5 antigen-like member of the scavenger receptor, cysteine-rich superfamily fibrinogen, albumin, and complement factor H (CFH). TRX-1 inhibited the alternative pathway C3 convertase, and its effect was additive with CFH.
These findings show that TRX-1 interacts with CFH, regulates complement activity, and inhibits CNV, suggesting novel preventive and interventional therapeutic strategies for AMD.
研究硫氧还蛋白-1（TRX-1），一种具有多种氧化还原相关作用的内源性蛋白质，在脉络膜新生血管（CNV）形成中的作用。
通过对野生型和过表达人TRX-1的转基因小鼠（TRX-1 Tg）进行眼底激光光凝诱导CNV。给小鼠腹腔注射TRX-1、突变型TRX或赋形剂。通过凝集素染色评估CNV的发生率。通过免疫组织化学和蛋白质印迹法测定激光损伤后的白细胞募集和C3b沉积。此外，使用与突变型TRX-1偶联的柱通过二维凝胶电泳从人血浆中分离TRX-1相关蛋白，并通过质谱和蛋白质组学分析进行鉴定。通过液相法测定补体激活。
与野生型小鼠（85.7%）相比，TRX-1 Tg小鼠（56.1%）和用TRX-1处理的C57B/6小鼠（46.7%）中激光诱导的CNV发生率降低，但突变型TRX-1处理的小鼠（79.2%）中未降低。此外，TRX-1处理的小鼠中白细胞募集受到抑制；这些小鼠和TRX-1 Tg小鼠中C3b沉积减少。在人血浆中，鉴定出与TRX-1相关的五种蛋白质为载脂蛋白A-I、清道夫受体富含半胱氨酸的超家族纤维蛋白原的CD5抗原样成员、白蛋白和补体因子H（CFH）。TRX-1抑制替代途径C3转化酶，其作用与CFH相加。
这些发现表明TRX-1与CFH相互作用，调节补体活性，并抑制CNV，提示年龄相关性黄斑变性的新型预防和介入治疗策略。
