Department of Ophthalmology, The First Affiliated Hospital, Xi'an Medical University, Xi'an, China.
Department of Ophthalmology, Xi'an Ninth Hospital Affiliated to Medical College of Xi'an Jiaotong University, Xi'an, China.
J Biochem Mol Toxicol. 2020 Nov;34(11):e22572. doi: 10.1002/jbt.22572. Epub 2020 Jul 7.
Angiogenic factor with G patch and FHA domains 1 (AGGF1) has strong proangiogenic effects on embryonic vascular development and angiogenesis in disease; however, its role in retinopathy has not been elucidated. Retinopathy of prematurity is a serious retinal disorder of premature infants, which is caused by the arrest of immature retinal vascular growth under hyperoxia. This study aims to investigate the effects of AGGF1 on retinal vascular endothelial cells under hyperoxia and the association with autophagy by using rhesus macaque choroid-retinal endothelial (RF/6A) cells. Western blot analysis and immunofluorescence staining were used to detect the expression of AGGF1 in RF/6A cells. Cell Counting Kit-8, flow cytometry, and transwell and matrigel assays were applied to detect the vitality, apoptosis, migration, and tube formation of RF/6A cells, respectively. Western blot analysis was then used to detect the expression of autophagy markers LC3 and Beclin-1, and mCherry-GFP-LC3 adenovirus was used to detect autophagy flux in RF/6A cells. Under hyperoxia, the expression of AGGF1 in RF/6A cells decreased compared with the control. Cell vitality, migration, and tube formation decreased, and apoptosis of RF/6A cells increased under hyperoxia, and these effects of hyperoxia were attenuated by AGGF1. The protein expressions of LC3 and Beclin-1 increased in RF/6A cells and autophagy flux enhanced under hyperoxia. AGGF1 reduced the expression of LC3 and Beclin-1 as well as the autophagy flux stimulated by hyperoxia. The results clearly showed that exogenous AGGF1 can protect retinal vascular endothelial cells and promote angiogenesis under hyperoxia, in which the expression of AGGF1 was inhibited. Inhibition of autophagy by AGGF1 may be one of the mechanisms involved.
血管生成因子 G 补丁和 FHA 结构域 1（AGGF1）在胚胎血管发育和疾病中的血管生成中具有很强的促血管生成作用；然而，其在视网膜病变中的作用尚未阐明。早产儿视网膜病变是一种严重的早产儿视网膜疾病，是由高氧下未成熟视网膜血管生长停滞引起的。本研究旨在使用恒河猴脉络膜视网膜内皮（RF/6A）细胞，研究 AGGF1 在高氧下对视网膜血管内皮细胞的影响及其与自噬的关系。Western blot 分析和免疫荧光染色用于检测 RF/6A 细胞中 AGGF1 的表达。细胞计数试剂盒-8、流式细胞术、Transwell 和 Matrigel 测定分别用于检测 RF/6A 细胞的活力、凋亡、迁移和管形成。然后用 Western blot 分析检测自噬标记物 LC3 和 Beclin-1 的表达，并用 mCherry-GFP-LC3 腺病毒检测 RF/6A 细胞中的自噬通量。在高氧条件下，与对照组相比，RF/6A 细胞中 AGGF1 的表达减少。高氧条件下 RF/6A 细胞活力、迁移和管形成减少，细胞凋亡增加，而 AGGF1 可减弱高氧的这些作用。在 RF/6A 细胞中，LC3 和 Beclin-1 的蛋白表达增加，自噬通量增强，而高氧刺激的自噬通量增强。结果清楚地表明，外源性 AGGF1 可在高氧下保护视网膜血管内皮细胞并促进血管生成，而 AGGF1 的表达受到抑制。AGGF1 抑制自噬可能是其参与的机制之一。
