Wu Ming-Xing, Zhou Yu, Zhou Xi-Yuan, Xu Yan
Department of Ophthalmology, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.
Int J Ophthalmol. 2022 Jun 18;15(6):876-885. doi: 10.18240/ijo.2022.06.04. eCollection 2022.
To investigate whether ultrasound-targeted cationic microbubbles (CMBs) destruction could deliver endostatin-green fluorescent protein (GFP) plasmids efficiently to the human retinal endothelial cells (HRECs) and inhibit retinal neovascularization in mice.
CMBs were prepared and the presentation of GFP reporter was confirmed by flow cytometry and laser confocal microscopy. Experiments assessing HRECs migration and vascular formation were performed to evaluate gene therapy's efficiency A mouse model of oxygen-induced retinopathy was employed and the expression of Bcl-xl, Bcl-2, vascular endothelial growth factor (VEGF) and endostatin in the retina of mice were determined by Western blotting and quantitative polymerase chain reaction (qPCR). The expression of endostatin-GFP in the retina was examined by laser confocal microscopy at 5, 14, and 28d after treatment.
The gene expression of endostatin was the highest in the group of the CMBs. Besides, the inhibition and antiangiogenesis effect of the migration and development of HRECs were improved following treatment with CMBs compared with the other groups . , retinal neovascularization was significantly inhibited and the fluorescence intensity of endostatin-GFP in the mouse retina was importantly higher in the group of CMBs than that in other groups.
The research illustrates ultrasound-targeted CMBs destruction possessed distinct effect on the inhibition of the vascular formation and the development of retinal neovascularization both and .
研究超声靶向破坏阳离子微泡(CMBs)能否将内皮抑素-绿色荧光蛋白(GFP)质粒有效递送至人视网膜内皮细胞(HRECs),并抑制小鼠视网膜新生血管形成。
制备CMBs,通过流式细胞术和激光共聚焦显微镜确认GFP报告基因的表达。进行评估HRECs迁移和血管形成的实验以评价基因治疗的效果。采用氧诱导视网膜病变小鼠模型,通过蛋白质免疫印迹法和定量聚合酶链反应(qPCR)检测小鼠视网膜中Bcl-xl、Bcl-2、血管内皮生长因子(VEGF)和内皮抑素的表达。在治疗后第5、14和28天,通过激光共聚焦显微镜检查视网膜中内皮抑素-GFP的表达。
内皮抑素基因表达在CMBs组中最高。此外,与其他组相比,用CMBs处理后HRECs迁移和发育的抑制及抗血管生成作用得到改善。CMBs组小鼠视网膜新生血管形成明显受到抑制,且内皮抑素-GFP在小鼠视网膜中的荧光强度显著高于其他组。
该研究表明超声靶向破坏CMBs对抑制血管形成和视网膜新生血管发育均具有显著作用。