Abstract

Introduction

To explore the effect and mechanism of APRPG-modified nanoliposomes loaded with miR-146a-5p inhibitor (ANL-miR-146a-5p inhibitor) on endothelial cell proliferation, migration, tube formation, and choroidal neovascularization (CNV) in mice.

Methods

ANL-miR-146a-5p inhibitors were generated by thin film hydration; in vitro, endothelial cell uptake experiment was used to detect the targeting effect of ANL-miR-146a-5p inhibitor; endothelial cells proliferation, migration, and tube formation were detected, respectively, by CCK8 assay, scratch assay, and Matrigel tube formation assay. In vivo, the mice CNV models were established by 810 nm laser photocoagulation. Mice choroidal flatmounts were performed to detect the volume of CNV after intravitreal injection of L-miR-146a-5p inhibitor, ANL-miR-146a-5p inhibitor, or normal saline; the vascular endothelial growth factor (VEGF) expression of mice choroidal tissue was detected by ELISA; HE section and electrophysiology (ERG) were performed to check the toxicity of ANL-miR-146a-5p inhibitor on mice retina.

Results

ANL are targeted to endothelial cells and are more targeted in inflammatory environment. At the same concentration, ANL-miR-146a-5p inhibitor’s ability to inhibit endothelial cell proliferation, migration, tube formation, CNV formation, and VEGF expression is better than L-miR-146a-5p inhibitor (p < 0.05); ANL-miR-146a-5p inhibitor had no toxicity on the structure of mouse retina; ANL-miR-146a-5p inhibitor had no toxicity on the a-wave and b-wave amplitudes and b-wave implicit times (p > 0.05).

Conclusions

ANL-miR-146a-5p inhibitor can more effectively down-regulate the expression level of VEGF through its targeting to endothelial cells, thereby more effectively inhibiting endothelial cell proliferation, migration, tube formation, and mice CNV formation.

中文翻译：

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载有miR-146a-5p抑制剂的APRPG修饰的纳米脂质体通过靶向内皮细胞来抑制脉络膜新血管形成。

摘要

介绍

探讨载有miR-146a-5p抑制剂（ANL-miR-146a-5p抑制剂）的APRPG修饰的纳米脂质体对小鼠内皮细胞增殖，迁移，管形成和脉络膜新血管形成（CNV）的作用和机制。

方法

ANL-miR-146a-5p抑制剂是通过薄膜水合作用产生的。在体外，通过内皮细胞摄取实验来检测ANL-miR-146a-5p抑制剂的靶向作用。通过CCK8测定，刮擦测定和Matrigel管形成测定分别检测了内皮细胞的增殖，迁移和管形成。在体内，通过810nm激光光凝建立小鼠CNV模型。玻璃体腔内注射L-miR-146a-5p抑制剂，ANL-miR-146a-5p抑制剂或生理盐水后，进行小鼠脉络膜平置术以检测CNV的体积；ELISA法检测小鼠脉络膜组织中血管内皮生长因子的表达。进行HE切片和电生理学（ERG）以检查ANL-miR-146a-5p抑制剂对小鼠视网膜的毒性。

结果

ANL靶向内皮细胞，在炎症环境中更具靶向性。在相同浓度下，ANL-miR-146a-5p抑制剂抑制内皮细胞增殖，迁移，管形成，CNV形成和VEGF表达的能力优于L-miR-146a-5p抑制剂（p  <0.05）；ANL-miR-146a-5p抑制剂对小鼠视网膜无毒性；ANL-miR-146a-5p抑制剂对a波和b波振幅以及b波隐含时间无毒性（p  > 0.05）。

结论

ANL-miR-146a-5p抑制剂可通过靶向内皮细胞更有效地下调VEGF的表达水平，从而更有效地抑制内皮细胞的增殖，迁移，管形成和小鼠CNV形成。