Department of Ophthalmology and Visual Science, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan.
Ophthalmologica. 2013;230(4):215-21. doi: 10.1159/000354092. Epub 2013 Sep 25.
Previous studies have shown that small interfering RNAs (siRNAs) could suppress angiogenesis via stimulation of toll-like receptor-3 (TLR3). The purpose of this study was to determine the efficacy of atelocollagen to deliver siRNA without TLR3 stimulation in the laser-induced choroidal neovascularization (CNV) model.
CNV was induced by laser injury in C57BL/6J mice and volumes were measured 7 days later. Nontargeted siRNA, 21-nucleotide (nt) siRNA-Luc (Luciferase) and 21-nt siRNA-Vegfa were injected into the vitreous following injury. Atelocollagen was incubated with naked 21-nt siRNAs before injection. To block TLR3 endosomal activity, chloroquine was injected intravitreously after laser injury.
The mean CNV volumes were significantly smaller in the naked siRNA-Luc, naked siRNA-Vegfa, or siRNA-Vegfa/atelocollagen complex compared with PBS, atelocollagen or siRNA-Luc/atelocollagen complex-injected mice (p < 0.05).
These findings demonstrate that atelocollagen may deliver siRNA without nonspecific TLR3 stimulation in the murine laser-CNV model.
先前的研究表明，小干扰 RNA（siRNA）可以通过刺激 Toll 样受体 3（TLR3）来抑制血管生成。本研究旨在确定无 TLR3 刺激的 ATelocollagen 递送 siRNA 在激光诱导脉络膜新生血管（CNV）模型中的疗效。
在 C57BL/6J 小鼠中通过激光损伤诱导 CNV，并在 7 天后测量体积。非靶向 siRNA、21 个核苷酸（nt）siRNA-Luc（荧光素酶）和 21-nt siRNA-Vegfa 在损伤后注入玻璃体。ATelocollagen 在注射前与裸 21-nt siRNA 孵育。为了阻断 TLR3 内体活性，在激光损伤后向玻璃体内注射氯喹。
与 PBS、ATelocollagen 或 siRNA-Luc/ATelocollagen 复合物注射小鼠相比，裸 siRNA-Luc、裸 siRNA-Vegfa 或 siRNA-Vegfa/ATelocollagen 复合物注射小鼠的平均 CNV 体积明显更小（p<0.05）。
这些发现表明，ATelocollagen 可能在小鼠激光-CNV 模型中无需非特异性 TLR3 刺激即可递送 siRNA。
