Activation of glial cells induces proinflammatory properties in brain capillary endothelial cells in vitro

Abstract Neurodegenerative diseases are often accompanied by neuroinflammation and impairment of the blood-brain barrier (BBB) mediated by activated glial cells through their release of proinflammatory molecules. To study the effects of glial cells on mouse brain endothelial cells (mBECs), we develo...

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Main Authors: Annette Burkhart, Steinunn Sara Helgudóttir, Yahye A. Mahamed, Mikkel B. Fruergaard, Julie N. Holm-Jacobsen, Hulda Haraldsdóttir, Sara E. Dahl, Freja Pretzmann, Lisa Greve Routhe, Kate Lambertsen, Torben Moos, Maj Schneider Thomsen
Format: Article
Language:English
Published: Nature Portfolio 2024-11-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-024-78204-w
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author Annette Burkhart
Steinunn Sara Helgudóttir
Yahye A. Mahamed
Mikkel B. Fruergaard
Julie N. Holm-Jacobsen
Hulda Haraldsdóttir
Sara E. Dahl
Freja Pretzmann
Lisa Greve Routhe
Kate Lambertsen
Torben Moos
Maj Schneider Thomsen
author_facet Annette Burkhart
Steinunn Sara Helgudóttir
Yahye A. Mahamed
Mikkel B. Fruergaard
Julie N. Holm-Jacobsen
Hulda Haraldsdóttir
Sara E. Dahl
Freja Pretzmann
Lisa Greve Routhe
Kate Lambertsen
Torben Moos
Maj Schneider Thomsen
author_sort Annette Burkhart
collection DOAJ
description Abstract Neurodegenerative diseases are often accompanied by neuroinflammation and impairment of the blood-brain barrier (BBB) mediated by activated glial cells through their release of proinflammatory molecules. To study the effects of glial cells on mouse brain endothelial cells (mBECs), we developed an in vitro BBB model with inflammation by preactivating mixed glial cells (MGCs) with lipopolysaccharide (LPS) before co-culturing with mBECs to study the influence of molecules released by activated MGCs. The response of the mBECs to activated MGCs was compared to direct stimulation with LPS. The cytokine profile of activated MGCs was analyzed together with their effects on the mBEC’s integrity, expression of tight junction proteins, adhesion molecules, and BBB-specific transport proteins. Stimulation of MGCs significantly upregulated mRNA expression and secretion of several pro-inflammatory cytokines. Co-culturing mBECs with pre-stimulated MGCs significantly affected the barrier integrity of mBECs similar to direct stimulation with LPS. The gene expression levels of tight junction proteins were unaltered, but tight junction proteins revealed rearrangements with respect to subcellular distribution. Compared to direct stimulation with LPS, the expression of cell-adhesion molecules was significantly increased when mBECs were co-cultured with prestimulated MGCs and thus pre-activating MGCs transforms mBECs into a proinflammatory phenotype.
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spelling doaj-art-77b46eebf90b409ab47df3068dca57dd2025-08-20T02:50:07ZengNature PortfolioScientific Reports2045-23222024-11-0114111610.1038/s41598-024-78204-wActivation of glial cells induces proinflammatory properties in brain capillary endothelial cells in vitroAnnette Burkhart0Steinunn Sara Helgudóttir1Yahye A. Mahamed2Mikkel B. Fruergaard3Julie N. Holm-Jacobsen4Hulda Haraldsdóttir5Sara E. Dahl6Freja Pretzmann7Lisa Greve Routhe8Kate Lambertsen9Torben Moos10Maj Schneider Thomsen11Neurobiology Research and Drug Delivery, Department of Health Science and Technology, Aalborg UniversityNeurobiology Research and Drug Delivery, Department of Health Science and Technology, Aalborg UniversityNeurobiology Research and Drug Delivery, Department of Health Science and Technology, Aalborg UniversityNeurobiology Research and Drug Delivery, Department of Health Science and Technology, Aalborg UniversityNeurobiology Research and Drug Delivery, Department of Health Science and Technology, Aalborg UniversityNeurobiology Research and Drug Delivery, Department of Health Science and Technology, Aalborg UniversityNeurobiology Research and Drug Delivery, Department of Health Science and Technology, Aalborg UniversityNeurobiology Research and Drug Delivery, Department of Health Science and Technology, Aalborg UniversityNeurobiology Research and Drug Delivery, Department of Health Science and Technology, Aalborg UniversityNeurobiology Research, Department of Molecular Medicine, University of Southern DenmarkNeurobiology Research and Drug Delivery, Department of Health Science and Technology, Aalborg UniversityNeurobiology Research and Drug Delivery, Department of Health Science and Technology, Aalborg UniversityAbstract Neurodegenerative diseases are often accompanied by neuroinflammation and impairment of the blood-brain barrier (BBB) mediated by activated glial cells through their release of proinflammatory molecules. To study the effects of glial cells on mouse brain endothelial cells (mBECs), we developed an in vitro BBB model with inflammation by preactivating mixed glial cells (MGCs) with lipopolysaccharide (LPS) before co-culturing with mBECs to study the influence of molecules released by activated MGCs. The response of the mBECs to activated MGCs was compared to direct stimulation with LPS. The cytokine profile of activated MGCs was analyzed together with their effects on the mBEC’s integrity, expression of tight junction proteins, adhesion molecules, and BBB-specific transport proteins. Stimulation of MGCs significantly upregulated mRNA expression and secretion of several pro-inflammatory cytokines. Co-culturing mBECs with pre-stimulated MGCs significantly affected the barrier integrity of mBECs similar to direct stimulation with LPS. The gene expression levels of tight junction proteins were unaltered, but tight junction proteins revealed rearrangements with respect to subcellular distribution. Compared to direct stimulation with LPS, the expression of cell-adhesion molecules was significantly increased when mBECs were co-cultured with prestimulated MGCs and thus pre-activating MGCs transforms mBECs into a proinflammatory phenotype.https://doi.org/10.1038/s41598-024-78204-wBlood-brain barrierNeuroinflammationIn-vitro BBB modelGlial cellsLipopolysaccharide
spellingShingle Annette Burkhart
Steinunn Sara Helgudóttir
Yahye A. Mahamed
Mikkel B. Fruergaard
Julie N. Holm-Jacobsen
Hulda Haraldsdóttir
Sara E. Dahl
Freja Pretzmann
Lisa Greve Routhe
Kate Lambertsen
Torben Moos
Maj Schneider Thomsen
Activation of glial cells induces proinflammatory properties in brain capillary endothelial cells in vitro
Scientific Reports
Blood-brain barrier
Neuroinflammation
In-vitro BBB model
Glial cells
Lipopolysaccharide
title Activation of glial cells induces proinflammatory properties in brain capillary endothelial cells in vitro
title_full Activation of glial cells induces proinflammatory properties in brain capillary endothelial cells in vitro
title_fullStr Activation of glial cells induces proinflammatory properties in brain capillary endothelial cells in vitro
title_full_unstemmed Activation of glial cells induces proinflammatory properties in brain capillary endothelial cells in vitro
title_short Activation of glial cells induces proinflammatory properties in brain capillary endothelial cells in vitro
title_sort activation of glial cells induces proinflammatory properties in brain capillary endothelial cells in vitro
topic Blood-brain barrier
Neuroinflammation
In-vitro BBB model
Glial cells
Lipopolysaccharide
url https://doi.org/10.1038/s41598-024-78204-w
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