Zhang Yue, Liu Xiao-Jing, Zhai Xin-Ran, Yao Yao, Shao Bin, Zhen Yu-Han, Zhang Xin, Xiao Zhe, Wang Li-Fang, Zhang Ming-Lian, Chen Zhi-Min
Hebei Eye Hospital, Provincial Key Laboratory of Ophthalmology, Hebei Provincial Clinical Research Center for Eye Diseases, Xingtai 054001, Hebei Province, China.
Department of Ophthalmology, Hebei Medical University, Shijiazhuang 050011, Hebei Province, China.
Int J Ophthalmol. 2024 Aug 18;17(8):1403-1410. doi: 10.18240/ijo.2024.08.03. eCollection 2024.
To investigate the effects of fibrillin-1 (FBN1) deletion on the integrity of retina-blood barrier function and the apoptosis of vascular endothelial cells under diabetic conditions.
Streptozotocin (STZ)-induced diabetic mice were used to simulate the diabetic conditions of diabetic retinopathy (DR) patients, and FBN1 expression was detected in retinas from STZ-diabetic mice and controls. In the Gene Expression Omnibus (GEO) database, the GSE60436 dataset was selected to analyze FBN1 expressions in fibrovascular membranes from DR patients. Using lentivirus to knock down FBN1 levels, vascular leakage and endothelial barrier integrity were detected by Evans blue vascular permeability assay, fluorescein fundus angiography (FFA) and immunofluorescence labeled with tight junction marker . High glucose-induced monkey retinal vascular endothelial cells (RF/6A) were used to investigate effects of FBN1 on the cells . The vascular endothelial barrier integrity and apoptosis were detected by trans-endothelial electrical resistance (TEER) assay and flow cytometry, respectively.
FBN1 mRNA expression was increased in retinas of STZ-induced diabetic mice and fibrovascular membranes of DR patients (GSE60436 datasets) using RNA-seq approach. Besides, knocking down of FBN1 by lentivirus intravitreal injection significantly inhibited the vascular leakage compared to STZ-DR group by Evans blue vascular permeability assay and FFA detection. Expressions of tight junction markers in STZ-DR mouse retinas were lower than those in the control group, and knocking down of FBN1 increased the tight junction levels. , 30 mmol/L glucose could significantly inhibit viability of RF/6A cells, and FBN1 mRNA expression was increased under 30 mmol/L glucose stimulation. Down-regulation of FBN1 reduced high glucose (HG)-stimulated retinal microvascular endothelial cell permeability, increased TEER, and inhibited RF/6A cell apoptosis .
The expression level of FBN1 increases in retinas and vascular endothelial cells under diabetic conditions. Down-regulation of FBN1 protects the retina of early diabetic rats from retina-blood barrier damage, reduce vascular leakage, cell apoptosis, and maintain vascular endothelial cell barrier function.
研究糖尿病条件下原纤维蛋白-1（FBN1）缺失对视网膜-血屏障功能完整性及血管内皮细胞凋亡的影响。
用链脲佐菌素（STZ）诱导的糖尿病小鼠模拟糖尿病视网膜病变（DR）患者的糖尿病状态，检测STZ诱导的糖尿病小鼠和对照组视网膜中FBN1的表达。在基因表达综合数据库（GEO）中，选择GSE60436数据集分析DR患者纤维血管膜中FBN1的表达。使用慢病毒降低FBN1水平，通过伊文思蓝血管通透性试验、荧光素眼底血管造影（FFA）和紧密连接标志物免疫荧光检测血管渗漏和内皮屏障完整性。用高糖诱导的猴视网膜血管内皮细胞（RF/6A）研究FBN1对细胞的影响。分别通过跨内皮电阻（TEER）试验和流式细胞术检测血管内皮屏障完整性和细胞凋亡。
采用RNA测序方法，STZ诱导的糖尿病小鼠视网膜和DR患者纤维血管膜中FBN1 mRNA表达增加。此外，通过慢病毒玻璃体内注射降低FBN1水平，与STZ-DR组相比，伊文思蓝血管通透性试验和FFA检测显示血管渗漏明显受到抑制。STZ-DR小鼠视网膜中紧密连接标志物的表达低于对照组，降低FBN1水平可增加紧密连接水平。30 mmol/L葡萄糖可显著抑制RF/6A细胞活力，在30 mmol/L葡萄糖刺激下FBN1 mRNA表达增加。下调FBN1可降低高糖（HG）刺激的视网膜微血管内皮细胞通透性，增加TEER，并抑制RF/6A细胞凋亡。
糖尿病条件下视网膜和血管内皮细胞中FBN1表达水平升高。下调FBN1可保护早期糖尿病大鼠视网膜免受视网膜-血屏障损伤，减少血管渗漏、细胞凋亡，并维持血管内皮细胞屏障功能。
