Astrocytes and pericytes attenuate severely injured patient plasma mediated expression of tight junction proteins in endothelial cells.
Astrocytes and pericytes attenuate severely injured patient plasma mediated expression of tight junction proteins in endothelial cells.
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DOI：
10.1371/journal.pone.0270817
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发表时间：
2022
期刊：
影响因子：
3.7
通讯作者：
中科院分区：
文献类型：
--
作者：
血脑屏障（BBB）破坏是脑损伤的继发性形式，其机制尚未完全阐明。现有研究表明，内皮细胞之间的紧密连接蛋白的分解是损伤后BBB通透性增加的主要驱动因素，并且神经血管单元的原代细胞（内皮细胞、星形胶质细胞和周细胞）之间的细胞间信号传导有助于紧密连接恢复。为了阐述这一研究主体，我们分析了严重损伤的患者血浆对单一培养中的每种细胞类型的影响，并一起在三培养模型中分析了紧密连接蛋白Claudins 3和5（CLDN 3，CLDN 5）和封闭带1（ZO-1）的转录和翻译表达。进行条件培养基转移研究以阐明负责差异紧密连接表达的细胞类型。我们的数据显示，与5%人离体严重损伤患者血浆孵育足以在内皮细胞紧密连接mRNA和蛋白质表达中产生差异反应。单一培养物中的内皮细胞产生CLDN 3和CLDN 5 mRNA表达的显著增加（相对于对照分别增加3.98和3.51倍，p<0.01）和CLDN 5蛋白表达的显著增加（相对于对照变化2.58倍，p<0.01），而在三培养物中，这种增加减弱。我们的三培养模型和条件培养基实验表明，来自星形胶质细胞和周细胞的条件培养基以及星形胶质细胞、周细胞和内皮细胞的三培养足以减弱在与严重损伤的创伤血浆孵育后的内皮单培养物中观察到的紧密连接蛋白CLDN 3和CLDN 5的转录增加。该数据表明，来自星形胶质细胞和周细胞的抑制性分子信号通过紧密连接CLDN 5的转录和翻译下调而导致损伤后BBB的长期破坏。
Blood Brain Barrier (BBB) breakdown is a secondary form of brain injury which has yet to be fully elucidated mechanistically. Existing research suggests that breakdown of tight junction proteins between endothelial cells is a primary driver of increased BBB permeability following injury, and intercellular signaling between primary cells of the neurovascular unit: endothelial cells, astrocytes, and pericytes; contribute to tight junction restoration. To expound upon this body of research, we analyzed the effects of severely injured patient plasma on each of the cell types in monoculture and together in a triculture model for the transcriptional and translational expression of the tight junction proteins Claudins 3 and 5, (CLDN3, CLDN5) and Zona Occludens 1 (ZO-1). Conditioned media transfer studies were performed to illuminate the cell type responsible for differential tight junction expression. Our data show that incubation with 5% human ex vivo severely injured patient plasma is sufficient to produce a differential response in endothelial cell tight junction mRNA and protein expression. Endothelial cells in monoculture produced a significant increase of CLDN3 and CLDN5 mRNA expression, (3.98 and 3.51 fold increase vs. control respectively, p<0.01) and CLDN5 protein expression, (2.58 fold change vs. control, p<0.01), whereas in triculture, this increase was attenuated. Our triculture model and conditioned media experiments suggest that conditioned media from astrocytes and pericytes and a triculture of astrocytes, pericytes and endothelial cells are sufficient in attenuating the transcriptional increases of tight junction proteins CLDN3 and CLDN5 observed in endothelial monocultures following incubation with severely injured trauma plasma. This data suggests that inhibitory molecular signals from astrocytes and pericytes contributes to prolonged BBB breakdown following injury via tight junction transcriptional and translational downregulation of CLDN5.
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