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25 September 2019 : Laboratory Research  

Basic Fibroblast Growth Factor Reduces Permeability and Apoptosis of Human Brain Microvascular Endothelial Cells in Response to Oxygen and Glucose Deprivation Followed by Reoxygenation via the Fibroblast Growth Factor Receptor 1 (FGFR1)/ERK Pathway

Peng Chen12EF, Hongguang Zhang1D, Qingtao Zhang2F, Wei Zhou2F, Yongbing Deng2B, Xi Hu2CE, Lianyang Zhang1AG*

DOI: 10.12659/MSM.918626

Med Sci Monit 2019; 25:7191-7201

Abstract

BACKGROUND: Disruption of the blood–brain barrier (BBB) is a mechanism in the pathogenesis of traumatic brain injury. Basic fibroblast growth factor (bFGF) is expressed in angiogenesis, neurogenesis, and neuronal survival. This study aimed to investigate the role of bFGF in vitro in human brain microvascular endothelial cells (HBMECs) challenged by oxygen-glucose deprivation/reperfusion (OGD/R).

MATERIAL AND METHODS: HBMECs were cultured in glucose-free medium and an environment with <0.5% oxygen in an anaerobic chamber. Immunocytochemistry, Western blot, and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) were used to measure the protein and mRNA expression levels of bFGF, tight junction, adherens junction, apoptotic proteins, and matrix metalloproteinases (MMPs). The effects of bFGF on the viability of HBMECs was evaluated using the cell counting kit-8 (CCK-8) assay. Cell apoptosis was evaluated using the TUNEL assay, and endothelial permeability was quantified using a transwell migration assay with fluorescein isothiocyanate (FITC) conjugated with dextran. The effects of bFGF were evaluated following inhibition of fibroblast growth factor receptor 1 (FGFR1) with PD173074 and inhibition of ERK with PD98059.

RESULTS: Following OGD/R of HBMECs, bFGF significantly reduced cell permeability and apoptosis and significantly inhibited the down-regulation of the expressions of proteins associated with tight junctions, adherens junctions, apoptosis and matrix metalloproteinases (MMPs). The effects of bFGF were mediated by the activation of FGFR1 and ERK, as they were blocked by FGFR1 and ERK inhibitors.

CONCLUSIONS: Permeability and apoptosis of HBMECs challenged by OGD/R were reduced by bFGF by activation of the FGFR1 and the ERK pathway.

Keywords: Blood-Brain Barrier, Brain Injuries, Receptor, Fibroblast Growth Factor, Type 2, Brain, Capillary Permeability, endothelial cells, Fibroblast Growth Factor 2, Glucose, Oxygen, primary cell culture, Receptor, Fibroblast Growth Factor, Type 1

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750