RESUMO
AIM: 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. METHODS: 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 in vivo. High glucose-induced monkey retinal vascular endothelial cells (RF/6A) were used to investigate effects of FBN1 on the cells in vitro. The vascular endothelial barrier integrity and apoptosis were detected by trans-endothelial electrical resistance (TEER) assay and flow cytometry, respectively. RESULTS: 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. In vitro, 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 in vitro. CONCLUSION: 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.
RESUMO
Dissolved organic matter(DOM) is an important component of the organic matter in the environment. This study explored the characteristics of DOM. The components and spectral characteristics of rainwater DOM were measured using Ultraviolet-visible absorption spectrum(UV-vis) and three-dimensional fluorescence spectrum in the Hebei province. The results showed significant differences for DOM molecular weight in the northern, central, and southern region; the DOM of the northern region exhibited the lowest molecular weight. Two humic-like(C1 and C4), one protein-like(C2) and one fulvic-like(C3) components were identified by parallel factor(PARAFAC) analysis. Component C2 had a significant positive correlation with components C3 and C4(P<0.001), which may have similar sources. The C2(protein-like substances) accounted for the major proportion of DOM with the average abundance 40.59%. The DOM components based on PARAFAC exhibited significantly difference between city and country regions without spatial difference, which was consistent with the result of fluorescence regional integration(FRI). The protein-like substances based on FRI were the main components, which accounted for 61.59%-89.01%. Redundancy analysis(RDA) showed that total nitrogen, nitrate, and ammonia were the main environmental factors determining the distribution of DOM. Spectral indices presented a significant difference between city and country regions. High fluorescence(FI) and biological(BIX) and low humification(HIX) values showed that summer rain exhibited the strong autochthonous and low humification characteristics, and country region have stronger autochthonous characteristics than city region. The regression analysis demonstrates that the prediction accuracy of rainwater quality parameters in city region is high. From all the results, rainwater DOM, showing strong autogenous characteristics, exhibited significant differences between city and country regions without spatial difference in Hebei. Meanwhile, it can also provide technical support for rainwater nitrogen concentration prediction based on DOM components.