We are analyzing https://link.springer.com/article/10.1007/s00125-020-05299-x.

Title:
Erythropoietin protects the inner blood–retinal barrier by inhibiting microglia phagocytosis via Src/Akt/cofilin signalling in experimental diabetic retinopathy | Diabetologia
Description:
Aims/hypothesis Microglial activation in diabetic retinopathy and the protective effect of erythropoietin (EPO) have been extensively studied. However, the regulation of microglia in the retina and its relationship to inner blood–retinal barrier (iBRB) maintenance have not been fully characterised. In this study, we investigated the role of microglia in iBRB breakdown in diabetic retinopathy and the protective effects of EPO in this context. Methods Male Sprague Dawley rats were injected intraperitoneally with streptozotocin (STZ) to establish the experimental model of diabetes. At 2 h after STZ injection, the right and left eyes were injected intravitreally with EPO (16 mU/eye, 2 μl) and an equivalent volume of normal saline (NaCl 154 mmol/l), respectively. The rats were killed at 2 or 8 weeks after diabetes onset. Microglia activation was detected by ionised calcium binding adaptor molecule (IBA)-1 immunolabelling. Leakage of the iBRB was evaluated by albumin staining and FITC-dextran permeability assay. BV2 cells and primary rat microglia under hypoxic conditions were used to model microglial activation in diabetic retinopathy. Phagocytosis was examined by confocal microscopy in flat-mounted retina preparations and in microglia and endothelial cell cocultures. Protein levels of IBA-1, CD11b, complement component 1r (C1r), and Src/Akt/cofilin signalling pathway components were assessed by western blotting. Results In diabetic rat retinas, phagocytosis of endothelial cells by activated microglia was observed at 8 weeks, resulting in an increased number of acellular capillaries (increased by 426.5%) and albumin leakage. Under hypoxic conditions, activated microglia transmigrated to the opposite membrane of the transwell, where they disrupted the endothelial cell monolayer by engulfing endothelial cells. The activation and phagocytic activity of microglia was blocked by intravitreal injection of EPO. In vitro, IBA-1, CD11b and C1r protein levels were increased by 50.9%, 170.0% and 135.5%, respectively, by hypoxia, whereas the phosphorylated proteins of Src/Akt/cofilin signalling pathway components were decreased by 74.2%, 47.8% and 39.7%, respectively, compared with the control; EPO treatment abrogated these changes. Conclusions/interpretation In experimental diabetic retinopathy, activated microglia penetrate the basement membrane of the iBRB and engulf endothelial cells, leading to iBRB breakdown. EPO exerts a protective effect that preserves iBRB integrity via activation of Src/Akt/cofilin signalling in microglia, as demonstrated in vitro. These data support a causal role for activated microglia in iBRB breakdown and highlight the therapeutic potential of EPO for the treatment of diabetic retinopathy.
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Keywords {🔍}

microglia, cells, diabetic, epo, retinopathy, endothelial, article, pubmed, conditions, fig, google, scholar, hypoxic, activation, retinal, cell, activated, retinas, hypoxia, phagocytosis, diabetes, number, treatment, catalogue, microglial, ibrb, iba, vascular, cas, increased, group, normal, leakage, data, esm, retina, rats, zhang, shanghai, albumin, phagocytic, control, compared, rat, usa, srcaktcofilin, signalling, breakdown, protein, normoxic,

Topics {✒️}

src/akt/cofilin signalling pathway rac1-mediated signaling pathway src/akt/cofilin signalling src/akt/cofilin pathway src/pi3k/rac1 signaling advanced glycation end-products donkey anti-goat igg donkey anti-mouse igg twenty-year incidence follow donkey anti-rabbit igg retinal flat-mount preparations fluospheres carboxylate-modified microspheres high-magnification view erythropoietin-derived peptide protects guo-tong xu prepare retinal flat-mounts p-cofilin/actin ratios retinal flat-mounts confirmed decreased integrin-β1 expression related subjects improved visual acuity fitc-dextran permeability assay microglia-derived inflammatory factors rabbit anti-iba-1 antibody 60 mg/kg body weight full access src family activator akt activator sc79 blood–retinal barrier actin-related protein rm/hrmec coculture system 8-week-diabetic rat retinas including amadori-glycated albumin endothelial cell fragments p-akt/actin jingfa zhang blocking/permeabilisation solution composed privacy choices/manage cookies flat-mounted retina preparations streptozotocin-induced diabetic rats blood–ocular barrier blood–brain barrier [46] lower integrin-β1 retinal vascular regression endothelial cell cocultures blood-retina barrier electronic supplementary material reducing pericyte loss endothelial cell medium complement component 1r

Questions {❓}

Karlstetter M, Scholz R, Rutar M, Wong WT, Provis JM, Langmann T (2015) Retinal microglia: just bystander or target for therapy?

Schema {🗺️}

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      pageEnd:225
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      keywords:
         Diabetic retinopathy
         Erythropoietin
         Inner blood–retinal barrier
         Microglia
         Retinal vascular endothelial cell
         Internal Medicine
         Metabolic Diseases
         Human Physiology
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