牙周炎与各种全身性疾病有关，牙龈卟啉单胞菌外膜囊泡（Pg-OMV）的通过可能会促进牙周炎的发生。最近的几项研究表明 Pg-OMV 与神经炎症和神经变性之间存在联系，但确切的因果关系仍不清楚。本研究旨在利用体外模型研究这些关联的机制。分离的 Pg-OMV 通过形态、大小和牙龈蛋白酶活性进行表征。我们将 SH-SY5Y 神经母细胞瘤细胞和 BV-2 小胶质细胞暴露于不同浓度的 Pg-OMV。使用细胞形态学、3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四唑测定、酶联免疫吸附测定和蛋白质印迹分析来评估 Pg-OMV 诱导的细胞机制神经元细胞的神经毒性和小胶质细胞的炎症反应。暴露于 Pg-OMV 会引起 SH-SY5Y 细胞的神经毒性，细胞收缩、活力降低、凋亡途径激活和神经元分化标记物减少证明了这一点。 Gingipain 抑制减轻了这些影响，表明 gingipain 介导 Pg-OMV 诱导的 SH-SY5Y 细胞神经毒性。我们对神经炎症的研究表明，BV-2 细胞内吞 Pg-OMV 后，脂多糖 (LPS) 可以通过激活涉及磷酸化 AKT 和磷酸化 JNK 途径的途径来调节诱导型一氧化氮合酶和肿瘤坏死因子-α 的产生。我们的研究表明，Pg-OMV 内吞后，牙龈蛋白酶可诱导 SH-SY5Y 细胞产生神经毒性。此外，Pg-OMV 相关的 LPS 可以通过 BV-2 细胞中的 AKT 和 JNK 信号通路触发神经炎症。© 2024 中华民国牙科科学协会。 Elsevier B.V. 的出版服务

Periodontitis is associated with various systemic diseases, potentially facilitated by the passage of Porphyromonas gingivalis outer membrane vesicles (Pg-OMVs). Several recent studies have suggested a connection between Pg-OMVs and neuroinflammation and neurodegeneration, but the precise causal relationship remains unclear. This study aimed to investigate the mechanisms underlying these associations using in vitro models.Isolated Pg-OMVs were characterized by morphology, size, and gingipain activity. We exposed SH-SY5Y neuroblastoma cells and BV-2 microglial cells to various concentrations of Pg-OMVs. Cell morphology, a 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, an enzyme-linked immunosorbent assay, and Western blot analysis were used to evaluate the cellular mechanism underlying Pg-OMV-induced neurotoxicity in neuronal cells and inflammatory responses in microglial cells.Exposure to Pg-OMVs induced neurotoxicity in SH-SY5Y cells, as evidenced by cellular shrinkage, reduced viability, activation of apoptotic pathways, and diminished neuronal differentiation markers. Gingipain inhibition mitigated these effects, suggesting that gingipain mediates Pg-OMVs-induced neurotoxicity in SH-SY5Y cells. Our research on neuroinflammation suggests that upon endocytosis of Pg-OMVs by BV-2 cells, lipopolysaccharide (LPS) can modulate the production of inducible nitric oxide synthase and tumor necrosis factor-alpha by activating pathways that involve phosphorylated AKT and the phosphorylated JNK pathway.Our study demonstrated that following the endocytosis of Pg-OMVs, gingipain can induce neurotoxicity in SH-SY5Y cells. Furthermore, the Pg-OMVs-associated LPS can trigger neuroinflammation via AKT and JNK signaling pathways in BV-2 cells.© 2024 Association for Dental Sciences of the Republic of China. Publishing services by Elsevier B.V.