Glucocorticoids exert differential effects on the endothelium in an in vitro model of the blood–retinal barrier

Anne-Eva van der Wijk, Paul Canning, Rutger P. van Heijningen, Ilse M. C. Vogels, Cornelis J. F. van Noorden, Ingeborg Klaassen, Reinier O. Schlingemann

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Purpose: Glucocorticoids (GCs) are used as treatment in diabetic macular oedema, a condition caused by blood–retinal barrier (BRB) disruption. The proposed mechanisms by which GCs reduce macular oedema are indirect anti-inflammatory effects and inhibition of VEGF production, but direct effects on the BRB endothelium may be equally important. Here, we investigated direct effects of GCs on the endothelium to understand the specific pathways of GC action, to enable development of novel therapeutics lacking the adverse side-effects of the presently used GCs. Methods: Primary bovine retinal endothelial cells (BRECs) were grown on Transwell inserts and treated with hydrocortisone (HC), dexamethasone (Dex) or triamcinolone acetonide (TA). Molecular barrier integrity of the BRB was determined by mRNA and protein expression, and barrier function was assessed using permeability assays. In addition, we investigated whether TA was able to prevent barrier disruption after stimulation with VEGF or cytokines. Results: Treatment of BRECs with GCs resulted in upregulation of tight junction mRNA (claudin-5, occludin, ZO-1) and protein (claudin-5 and ZO-1). In functional assays, only TA strengthened the barrier function by reducing endothelial permeability. Moreover, TA was able to prevent cytokine-induced permeability in human retinal endothelial cells and VEGF-induced expression of plasmalemma vesicle-associated protein (PLVAP), a key player in VEGF-induced retinal vascular leakage. Conclusion: Glucocorticoids have differential effects in an experimental in vitro BRB model. TA is the most potent in improving barrier function, both at the molecular and functional levels, and TA prevents VEGF-induced expression of PLVAP.
Original languageEnglish
Pages (from-to)214-224
JournalActa Ophthalmologica
Volume97
Issue number2
DOIs
Publication statusPublished - 1 Mar 2019

Cite this

van der Wijk, A-E., Canning, P., van Heijningen, R. P., Vogels, I. M. C., van Noorden, C. J. F., Klaassen, I., & Schlingemann, R. O. (2019). Glucocorticoids exert differential effects on the endothelium in an in vitro model of the blood–retinal barrier. Acta Ophthalmologica, 97(2), 214-224. https://doi.org/10.1111/aos.13909
van der Wijk, Anne-Eva ; Canning, Paul ; van Heijningen, Rutger P. ; Vogels, Ilse M. C. ; van Noorden, Cornelis J. F. ; Klaassen, Ingeborg ; Schlingemann, Reinier O. / Glucocorticoids exert differential effects on the endothelium in an in vitro model of the blood–retinal barrier. In: Acta Ophthalmologica. 2019 ; Vol. 97, No. 2. pp. 214-224.
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title = "Glucocorticoids exert differential effects on the endothelium in an in vitro model of the blood–retinal barrier",
abstract = "Purpose: Glucocorticoids (GCs) are used as treatment in diabetic macular oedema, a condition caused by blood–retinal barrier (BRB) disruption. The proposed mechanisms by which GCs reduce macular oedema are indirect anti-inflammatory effects and inhibition of VEGF production, but direct effects on the BRB endothelium may be equally important. Here, we investigated direct effects of GCs on the endothelium to understand the specific pathways of GC action, to enable development of novel therapeutics lacking the adverse side-effects of the presently used GCs. Methods: Primary bovine retinal endothelial cells (BRECs) were grown on Transwell inserts and treated with hydrocortisone (HC), dexamethasone (Dex) or triamcinolone acetonide (TA). Molecular barrier integrity of the BRB was determined by mRNA and protein expression, and barrier function was assessed using permeability assays. In addition, we investigated whether TA was able to prevent barrier disruption after stimulation with VEGF or cytokines. Results: Treatment of BRECs with GCs resulted in upregulation of tight junction mRNA (claudin-5, occludin, ZO-1) and protein (claudin-5 and ZO-1). In functional assays, only TA strengthened the barrier function by reducing endothelial permeability. Moreover, TA was able to prevent cytokine-induced permeability in human retinal endothelial cells and VEGF-induced expression of plasmalemma vesicle-associated protein (PLVAP), a key player in VEGF-induced retinal vascular leakage. Conclusion: Glucocorticoids have differential effects in an experimental in vitro BRB model. TA is the most potent in improving barrier function, both at the molecular and functional levels, and TA prevents VEGF-induced expression of PLVAP.",
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van der Wijk, A-E, Canning, P, van Heijningen, RP, Vogels, IMC, van Noorden, CJF, Klaassen, I & Schlingemann, RO 2019, 'Glucocorticoids exert differential effects on the endothelium in an in vitro model of the blood–retinal barrier' Acta Ophthalmologica, vol. 97, no. 2, pp. 214-224. https://doi.org/10.1111/aos.13909

Glucocorticoids exert differential effects on the endothelium in an in vitro model of the blood–retinal barrier. / van der Wijk, Anne-Eva; Canning, Paul; van Heijningen, Rutger P.; Vogels, Ilse M. C.; van Noorden, Cornelis J. F.; Klaassen, Ingeborg; Schlingemann, Reinier O.

In: Acta Ophthalmologica, Vol. 97, No. 2, 01.03.2019, p. 214-224.

Research output: Contribution to journalArticleAcademicpeer-review

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T1 - Glucocorticoids exert differential effects on the endothelium in an in vitro model of the blood–retinal barrier

AU - van der Wijk, Anne-Eva

AU - Canning, Paul

AU - van Heijningen, Rutger P.

AU - Vogels, Ilse M. C.

AU - van Noorden, Cornelis J. F.

AU - Klaassen, Ingeborg

AU - Schlingemann, Reinier O.

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Y1 - 2019/3/1

N2 - Purpose: Glucocorticoids (GCs) are used as treatment in diabetic macular oedema, a condition caused by blood–retinal barrier (BRB) disruption. The proposed mechanisms by which GCs reduce macular oedema are indirect anti-inflammatory effects and inhibition of VEGF production, but direct effects on the BRB endothelium may be equally important. Here, we investigated direct effects of GCs on the endothelium to understand the specific pathways of GC action, to enable development of novel therapeutics lacking the adverse side-effects of the presently used GCs. Methods: Primary bovine retinal endothelial cells (BRECs) were grown on Transwell inserts and treated with hydrocortisone (HC), dexamethasone (Dex) or triamcinolone acetonide (TA). Molecular barrier integrity of the BRB was determined by mRNA and protein expression, and barrier function was assessed using permeability assays. In addition, we investigated whether TA was able to prevent barrier disruption after stimulation with VEGF or cytokines. Results: Treatment of BRECs with GCs resulted in upregulation of tight junction mRNA (claudin-5, occludin, ZO-1) and protein (claudin-5 and ZO-1). In functional assays, only TA strengthened the barrier function by reducing endothelial permeability. Moreover, TA was able to prevent cytokine-induced permeability in human retinal endothelial cells and VEGF-induced expression of plasmalemma vesicle-associated protein (PLVAP), a key player in VEGF-induced retinal vascular leakage. Conclusion: Glucocorticoids have differential effects in an experimental in vitro BRB model. TA is the most potent in improving barrier function, both at the molecular and functional levels, and TA prevents VEGF-induced expression of PLVAP.

AB - Purpose: Glucocorticoids (GCs) are used as treatment in diabetic macular oedema, a condition caused by blood–retinal barrier (BRB) disruption. The proposed mechanisms by which GCs reduce macular oedema are indirect anti-inflammatory effects and inhibition of VEGF production, but direct effects on the BRB endothelium may be equally important. Here, we investigated direct effects of GCs on the endothelium to understand the specific pathways of GC action, to enable development of novel therapeutics lacking the adverse side-effects of the presently used GCs. Methods: Primary bovine retinal endothelial cells (BRECs) were grown on Transwell inserts and treated with hydrocortisone (HC), dexamethasone (Dex) or triamcinolone acetonide (TA). Molecular barrier integrity of the BRB was determined by mRNA and protein expression, and barrier function was assessed using permeability assays. In addition, we investigated whether TA was able to prevent barrier disruption after stimulation with VEGF or cytokines. Results: Treatment of BRECs with GCs resulted in upregulation of tight junction mRNA (claudin-5, occludin, ZO-1) and protein (claudin-5 and ZO-1). In functional assays, only TA strengthened the barrier function by reducing endothelial permeability. Moreover, TA was able to prevent cytokine-induced permeability in human retinal endothelial cells and VEGF-induced expression of plasmalemma vesicle-associated protein (PLVAP), a key player in VEGF-induced retinal vascular leakage. Conclusion: Glucocorticoids have differential effects in an experimental in vitro BRB model. TA is the most potent in improving barrier function, both at the molecular and functional levels, and TA prevents VEGF-induced expression of PLVAP.

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van der Wijk A-E, Canning P, van Heijningen RP, Vogels IMC, van Noorden CJF, Klaassen I et al. Glucocorticoids exert differential effects on the endothelium in an in vitro model of the blood–retinal barrier. Acta Ophthalmologica. 2019 Mar 1;97(2):214-224. https://doi.org/10.1111/aos.13909