Cholesterol modification of SDF-1-specific siRNA enables therapeutic targeting of angiogenesis through Akt pathway inhibition
Cholesterol modification of SDF-1-specific siRNA enables therapeutic targeting of angiogenesis through Akt pathway inhibition
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胆固醇修饰 SDF-1 特异性 siRNA 可通过抑制 Akt 通路实现血管生成的治疗目标
DOI：
10.1016/j.exer.2019.03.006
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发表时间：
2019-07-01
影响因子：
3.4
通讯作者：
Lu, Linna
中科院分区：
文献类型：
--
作者：
Chen, Junzhao;Li, Fang;Lu, Linna
眼组织修复过程中的新生血管可导致严重的视轴视力丧失，因此是眼科医生相当关注的问题。在这里，我们介绍了一种胆固醇修饰的siRNA递送系统，靶向基质细胞衍生因子1（SDF-1），以治疗体内眼部血管生成。采用定量PCR（qPCR）分析大鼠内皮祖细胞（EPCs）和骨髓间充质干细胞（BMSCs）中SDF-1的表达。transwell法检测BMSC和HUVEC的迁移能力。集落形成实验检测chol-siSDF 1对HUVEC增殖的影响。在角膜碱烧伤模型中测试chol-siSDF 1的体内抗血管生成作用，并使用计算机成像分析系统测量角膜新生血管的面积。Western blot检测磷酸化Akt和总Akt蛋白水平。结果表明，大鼠EPCs和BMSCs均显示SDF-1 mRNA的高表达，而chol-siSDF-1可下调其表达。Chol-siSDF-1可显著抑制BMSC和HUVEC的迁移。此外，chol-siSDF 1还能抑制角膜碱烧伤模型中HUVEC的增殖，发挥显著的抗血管生成作用。chol-siSDF 1可能通过抑制Akt信号通路抑制肿瘤血管生成、增殖和转移。因此，靶向SDF-1的siRNA的胆固醇修饰显示出对迁移和血管生成的有效抑制，具有长得多的抑制作用持续时间。
Neovascularization during ocular tissue repair can cause severe visual loss in the optical axis and is therefore an issue of considerable concern to ophthalmologists. Here, we introduced a cholesterol-modified siRNA delivery system targeting stromal cell-derived factor 1 (SDF-1) to treat ocular angiogenesis in vivo. SDF-1 expression was analyzed in rat endothelial progenitor cells (EPCs) and bone marrow mesenchymal stem cells (BMSCs) using quantitative PCR (qPCR). Migration ability of BMSC and HUVEC were assessed through transwell assay. The proliferation effect of chol-siSDF1 on HUVEC was measured by colony formation assay. In vivo anti-angiogenic effects of chol-siSDF1 were tested in a cornea alkali burn model and the area of cornea neovascularization was measured using computer-imaging analysis system. Then phosphorylated Akt and total Akt protein levels were measured through western blot. Results turned out that rat EPCs and BMSCs showed high SDF-1 mRNA expression, which can be down-regulated by using chol-siSDF-1. Chol-siSDF-1 could significantly inhibit migration of BMSC and HUVEC. In addition, chol-siSDF1 also could inhibit HUVEC proliferation and exert a significant anti-angiogenic effect in corneal alkali burn model. As for the mechanism, chol-siSDF1 may inhibit the neovascularization, proliferation and metastasis through inhibiting the Akt signaling pathway. Thus, cholesterol modification of siRNA targeting SDF-1 displays an effective inhibition of migration and angiogenesis, with a much longer duration of inhibition effect.