Influences of advanced glycosylation end products on the inner blood-retinal barrier in a co-cultural cell model in vitro [post]

2020 unpublished
Advanced glycosylation end products (AGEs) are harmful factors that can damage the inner blood-retinal barrier (iBRB). However, their effects on iBRB co-culture models in vitro have not been reported. This study is to understand the interactive effects of different concentrations of AGEs at different time points on rat retinal microvascular endothelial cells (RMEC) and rat retinal Müller glial cell (RMGC) co-culture models. Methods : RMEC of Sprague-Dawley rat was isolated and cultured,
more » ... ed by anti-CD31 flow cytometry and immunocytometry with von Willebrand factor polyclonal antibody. Similarly, RMGC of Sprague-Dawley rat was identified by H&E staining, and immunohistochemical method with antibodies of Glial fibrillary acidic protein (GFAP) and glutamine synthetase (GS). The transepithelial electrical resistance (TEER) value was measured with the Millicell electrical resistance system to observe the leakage of the barrier. The Transwell cell for co-cultured RMEC with RMGC were used to construct an iBRB model and tested with the addition of AGEs at final concentrations of 50 mg/L and 100 mg/L, respectively, for 24 hours, 48 hours, and 72 hours. Results: AGEs in vitro iBRB model constructed by RMEC and RMGC co-culture led to the imbalance of VEGF and PEDF, and the permeability of the RMEC layer increased because TEER decreased in a doseand time-dependent manner. In the AGEs intervention the vascular endothelial growth factor (VEGF) was increased, while pigment epithelial derivative factor (PEDF) decreased, respectively, in a doseand time-dependent manner by enzyme-linked immunosorbent assay. Conclusions: The intervention with AGEs led to change of the RMEC layer transendothelial resistance and ratio of VEGF/PEDF. The iBRB in vitro model is a good tool to study the pathogenesis of retinal vascular diseases such as diabetic retinopathy and to evaluate the candidate drugs on the diseases. Background The blood-retinal barrier (BRB) is a special structure in the retina that regulates the exchange of substances inside and outside the blood vessels of the retina. Human and certain species of animals have dual blood supply systems: retinal blood vessels and choroidal blood vessels. The BRB corresponding to the two vascular systems are the inner blood-retina barrier (iBRB) and outer blood consent was obtained from themselves.
doi:10.21203/rs.2.23534/v1 fatcat:v3xgbfvrbnejjoghuefaejfinu