Department of Medical Microbiology and Nanobiomedical Engineering, Medical University of Białystok, Mickiewicza 2C, 15-222, Białystok, Poland.
Department of Biomedical Engineering, Rowan University, Glassboro, NJ, 08028, USA.
J Neuroinflammation. 2022 Nov 24;19(1):282. doi: 10.1186/s12974-022-02642-4.
Plasma gelsolin (pGSN) is an important part of the blood actin buffer that prevents negative consequences of possible F-actin deposition in the microcirculation and has various functions during host immune response. Recent reports reveal that severe COVID-19 correlates with reduced levels of pGSN. Therefore, using an in vitro system, we investigated whether pGSN could attenuate increased permeability of the blood-brain barrier (BBB) during its exposure to the portion of the SARS-CoV-2 spike protein containing the receptor binding domain (S1 subunit).
Two- and three-dimensional models of the human BBB were constructed using the human cerebral microvascular endothelial cell line hCMEC/D3 and exposed to physiologically relevant shear stress to mimic perfusion in the central nervous system (CNS). Trans-endothelial electrical resistance (TEER) as well as immunostaining and Western blotting of tight junction (TJ) proteins assessed barrier integrity in the presence of the SARS-CoV-2 spike protein and pGSN. The IncuCyte Live Imaging system evaluated the motility of the endothelial cells. Magnetic bead-based ELISA was used to determine cytokine secretion. Additionally, quantitative real-time PCR (qRT-PCR) revealed gene expression of proteins from signaling pathways that are associated with the immune response.
pGSN reversed S1-induced BBB permeability in both 2D and 3D BBB models in the presence of shear stress. BBB models exposed to pGSN also exhibited attenuated pro-inflammatory signaling pathways (PI3K, AKT, MAPK, NF-κB), reduced cytokine secretion (IL-6, IL-8, TNF-α), and increased expression of proteins that form intercellular TJ (ZO-1, occludin, claudin-5).
Due to its anti-inflammatory and protective effects on the brain endothelium, pGSN has the potential to be an alternative therapeutic target for patients with severe SARS-CoV-2 infection, especially those suffering neurological complications of COVID-19.
血浆凝胶蛋白（pGSN）是血液肌动蛋白缓冲液的重要组成部分，可防止肌动蛋白在微循环中沉积带来的负面后果，并在宿主免疫反应中发挥多种功能。最近的报告显示，严重的 COVID-19 与 pGSN 水平降低有关。因此，我们使用体外系统研究了 pGSN 是否可以减轻 SARS-CoV-2 刺突蛋白的受体结合域（S1 亚基）暴露于血脑屏障（BBB）通透性增加的情况。
使用人脑血管内皮细胞系 hCMEC/D3 构建了二维和三维的 BBB 模型，并施加生理相关的切应力以模拟中枢神经系统（CNS）中的灌注。跨内皮电阻（TEER）以及紧密连接（TJ）蛋白的免疫染色和 Western blot 检测在 SARS-CoV-2 刺突蛋白和 pGSN 存在的情况下，屏障的完整性。Incucyte 活细胞成像系统评估内皮细胞的迁移能力。磁珠酶联免疫吸附试验（ELISA）用于确定细胞因子的分泌。此外，定量实时 PCR（qRT-PCR）揭示了与免疫反应相关的信号通路蛋白的基因表达。
pGSN 在存在切应力的情况下，逆转了 S1 诱导的二维和三维 BBB 模型中的 BBB 通透性。暴露于 pGSN 的 BBB 模型还表现出减弱的促炎信号通路（PI3K、AKT、MAPK、NF-κB）、减少细胞因子分泌（IL-6、IL-8、TNF-α）和增加形成细胞间 TJ 的蛋白表达（ZO-1、occludin、claudin-5）。
由于其对大脑内皮的抗炎和保护作用，pGSN 有可能成为严重 SARS-CoV-2 感染患者的治疗靶点，特别是那些患有 COVID-19 神经系统并发症的患者。
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