State Key Laboratory of Trauma, Burns and Combined Injury, Trauma Center of People's Liberation Army (PLA), Daping Hospital, Army Medical University, Chongqing, China (mainland).
Department of Neurosurgery, Chongqing Emergency Medical Center, Chongqing, China (mainland).
Med Sci Monit. 2019 Sep 25;25:7191-7201. doi: 10.12659/MSM.918626.
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.
血脑屏障（BBB）的破坏是创伤性脑损伤发病机制中的一种机制。碱性成纤维细胞生长因子（bFGF）在血管生成、神经发生和神经元存活中表达。本研究旨在探讨 bFGF 在体外对氧葡萄糖剥夺/再灌注（OGD/R）挑战的人脑血管内皮细胞（HBMEC）中的作用。
将 HBMEC 在无糖培养基中培养，并在厌氧室中处于<0.5%氧气的环境中。免疫细胞化学、Western blot 和定量逆转录聚合酶链反应（qRT-PCR）用于测量 bFGF、紧密连接、黏附连接、凋亡蛋白和基质金属蛋白酶（MMPs）的蛋白和 mRNA 表达水平。使用细胞计数试剂盒-8（CCK-8）测定 bFGF 对 HBMEC 活力的影响。使用 TUNEL 测定评估细胞凋亡，使用 FITC 缀合的葡聚糖定量测定透膜迁移测定内皮通透性。用 PD173074 抑制成纤维细胞生长因子受体 1（FGFR1）和 PD98059 抑制 ERK 后评估 bFGF 的作用。
在 HBMEC 进行 OGD/R 后，bFGF 显著降低了细胞通透性和凋亡，并显著抑制了与紧密连接、黏附连接、凋亡和基质金属蛋白酶（MMPs）相关的蛋白表达下调。bFGF 的作用是通过激活 FGFR1 和 ERK 介导的，因为它们被 FGFR1 和 ERK 抑制剂阻断。
bFGF 通过激活 FGFR1 和 ERK 通路，减少了 OGD/R 挑战的 HBMEC 的通透性和凋亡。
