Neurobiology Research and Drug Delivery, Department of Health Science and Technology, Aalborg University, Selma Lagerlöfts Vej 249, 9260, Gistrup, Denmark.
Neurobiology Research, Department of Molecular Medicine, University of Southern Denmark, Campusvej 55, 5230, Odense M, Denmark.
Sci Rep. 2024 Nov 4;14(1):26580. doi: 10.1038/s41598-024-78204-w.
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.
神经退行性疾病通常伴随着神经炎症和血脑屏障（BBB）的损伤，这是由激活的神经胶质细胞通过释放促炎分子介导的。为了研究神经胶质细胞对小鼠脑内皮细胞（mBEC）的影响，我们通过用脂多糖（LPS）预先激活混合神经胶质细胞（MGC），然后与 mBEC 共培养来建立炎症体外 BBB 模型，以研究激活的 MGC 释放的分子的影响。将 mBEC 对激活的 MGC 的反应与 LPS 的直接刺激进行了比较。分析了激活的 MGC 的细胞因子谱及其对 mBEC 完整性、紧密连接蛋白表达、粘附分子和 BBB 特异性转运蛋白的影响。MGC 的刺激显著上调了几种促炎细胞因子的 mRNA 表达和分泌。与 LPS 的直接刺激相比，与预先刺激的 MGC 共培养 mBECs 显著影响 mBECs 的屏障完整性。紧密连接蛋白的基因表达水平没有改变，但紧密连接蛋白在亚细胞分布方面发生了重排。与 LPS 的直接刺激相比，当 mBECs 与预刺激的 MGC 共培养时，细胞粘附分子的表达显著增加，因此，预先激活 MGC 将 mBEC 转化为促炎表型。
