Silver nanoparticles inhibit VEGF-and IL-1β-induced vascular permeability via Src dependent pathway in porcine retinal endothelial cells

Abstract

The aim of this study is to determine the effects of silver nanoparticles (Ag-NP) on vascular endothelial growth factor (VEGF)-and interleukin-1 beta (IL-1β)-induced vascular permeability, and to detect the underlying signaling mechanisms involved in endothelial cells. Porcine retinal endothelial cells (PRECs) were exposed to VEGF, IL-1β and Ag-NP at different combinations and endothelial cell permeability was analyzed by measuring the flux of RITC-dextran across the PRECs monolayer. We found that VEGF and IL-1β increase flux of dextran across a PRECs monolayer, and Ag-NP block solute flux induced by both VEGF and IL-1β. To explore the signalling pathway involved VEGF- and IL-1β-induced endothelial alteration, PRECs were treated with Src inhibitor PP2 prior to VEGF and IL-1β treatment, and the effects were recorded. Further, to clarify the possible involvement of the Src pathways in endothelial cell permeability, plasmid encoding dominant negative(DN) and constitutively active(CA) form of Src kinases were transfected into PRECs, 24 h prior to VEGF and IL-1β exposure and the effects were recorded. Overexpression of DN Src blocked both VEGF-and IL-1β-induced permeability, while overexpression of CA Src rescues the inhibitory action of Ag-NP in the presence or absence of VEGF and IL-1β. Further, an in vitro kinase assay was performed to identify the presence of the Src phosphorylation at Y419. We report that VEGF and IL-1β-stimulate endothelial permeability via Src dependent pathway by increasing the Src phosphorylation and Ag-NP block the VEGF-and IL-1β-induced Src phosphorylation at Y419. These results demonstrate that Ag-NP may inhibit the VEGF-and IL-1β-induced permeability through inactivation of Src kinase pathway and this pathway may represent a potential therapeutic target to inhibit the ocular diseases such as diabetic retinopathy.

Epileptiform Activity Induces Vascular Remodeling and Zonula Occludens 1 Downregulation in Organotypic Hippocampal Cultures: Role of VEGF Signaling Pathways

“…We showed here that downregulation of ZO-1, an essential scaffold protein for tight junction assembly, follows activation of VEGFR-2 and Src pathway. We also showed that Src inhibition provided an effective protection of ZO-1 network after SLEs, confirming the major role of Src in ZO-1 downregulation, as was reported in models of ischemia and retinopathy (Sheikpranbabu et al, 2009;Takenaga et al, 2009). It is noteworthy that another Src inhibitor (CGP76030), tested on an in vivo model of epilepsy, was shown to reduce the proconvulsive effects of IL-1␤ , strengthening the interest in investigating Src signaling for epilepsy.…”

Antitumor activity of silver nanoparticles in Dalton’s lymphoma ascites tumor model

“…23 Silver nanoparticles inhibit vascular endothelial growth factor (VEGF)-induced angiogenesis in bovine retinal endothelial cells. 24 Similar studies have proven their inhibitory effect on vascular permeability induced by VEGF, interleukin (IL)-1β, 25 and advanced glycation end product 26 in retinal endothelial cells. The potent antiangiogenic and antipermeability effects of AgNPs, along with their ability to halt tumor progression in Pliss lymphosarcoma cells, have prompted the study of the antitumor effect of AgNPs in ascitic tumors.…”

Biofabrication of a novel biomolecule-assisted reduced graphene oxide: an excellent biocompatible nanomaterial

“…It could also favor the healing of corneal epithelial wounds. Previous studies demonstrated that low concentrations of silver nanoparticles rescued vascular endothelial growth factor and advanced glycation end-products induced vascular permeability through upregulation of ZO-1 and occludin 80,81 in porcine retinal endothelial cells. Our data are consistent with previous reports demonstrating that tight junction is important for proper cell function, which can be maintained by the treatment of cells with NAM-rGO.…”