Department of Neurology, Tianjin Nankai Hospital, Tianjin Medical University, Tianjin, 300052, China.
Department of Neurology, Tianjin Neurological Institute, Tianjin Medical University General Hospital, Tianjin, 300052, China.
J Neuroinflammation. 2024 Mar 25;21(1):73. doi: 10.1186/s12974-024-03057-z.
Guillain-Barré syndrome (GBS), a post-infectious, immune-mediated, acute demyelinating disease of the peripheral nerves and nerve roots, represents the most prevalent and severe acute paralyzing neuropathy. Purinergic P2X7 receptors (P2X7R) play a crucial role in central nervous system inflammation. However, little is known about their role in the immune-inflammatory response within the peripheral nervous system.
Initially, we assessed the expression of purinergic P2X7R in the peripheral blood of patients with GBS using flow cytometry and qRT-PCR. Next, we explored the expression of P2 X7R in CD4 T cells, CD8 T cells, and macrophages within the sciatic nerves and spleens of rats using immunofluorescence labeling and flow cytometry. The P2X7R antagonist brilliant blue G (BBG) was employed to examine its therapeutic impact on rats with experimental autoimmune neuritis (EAN) induced by immunization with the P0 peptide. We analyzed CD4 T cell differentiation in splenic mononuclear cells using flow cytometry, assessed Th17 cell differentiation in the sciatic nerve through immunofluorescence staining, and examined the expression of pro-inflammatory cytokine mRNA using RT-PCR. Additionally, we performed protein blotting to assess the expression of P2X7R and NLRP3-related inflammatory proteins within the sciatic nerve. Lastly, we utilized flow cytometry and immunofluorescence labeling to examine the expression of NLRP3 on CD4 T cells in rats with EAN.
P2X7R expression was elevated not only in the peripheral blood of patients with GBS but also in rats with EAN. In rats with EAN, inhibiting P2X7R with BBG alleviated neurological symptoms, reduced demyelination, decreased inflammatory cell infiltration of the peripheral nerves, and improved nerve conduction. BBG also limited the production of pro-inflammatory molecules, down-regulated the expression of P2X7R and NLRP3, and suppressed the differentiation of Th1 and Th17 cells, thus protecting against EAN. These effects collectively contribute to modifying the inflammatory environment and enhancing outcomes in EAN rats.
Suppression of P2X7R relieved EAN manifestation by regulating CD4 T cell differentiation and NLRP3 inflammasome activation. This finding underscores the potential significance of P2X7R as a target for anti-inflammatory treatments, advancing research and management of GBS.
吉兰-巴雷综合征（GBS）是一种感染后发生的、免疫介导的急性脱髓鞘性周围神经病，是最常见和最严重的急性麻痹性神经病。嘌呤能 P2X7 受体（P2X7R）在中枢神经系统炎症中发挥关键作用。然而，关于其在外周神经系统免疫炎症反应中的作用知之甚少。
最初，我们使用流式细胞术和 qRT-PCR 评估了 GBS 患者外周血中嘌呤能 P2X7R 的表达。接下来，我们使用免疫荧光标记和流式细胞术研究了 P2X7R 在大鼠坐骨神经和脾脏中的 CD4 T 细胞、CD8 T 细胞和巨噬细胞中的表达。使用嘌呤能 P2X7R 拮抗剂 brilliant blue G（BBG）来检查其对免疫接种 P0 肽诱导的实验性自身免疫性神经炎（EAN）大鼠的治疗作用。我们使用流式细胞术分析脾单核细胞中 CD4 T 细胞的分化，通过免疫荧光染色评估坐骨神经中 Th17 细胞的分化，并使用 RT-PCR 检查促炎细胞因子 mRNA 的表达。此外，我们进行蛋白质印迹以评估坐骨神经中 P2X7R 和 NLRP3 相关炎症蛋白的表达。最后，我们使用流式细胞术和免疫荧光标记检查 EAN 大鼠中 CD4 T 细胞上 NLRP3 的表达。
不仅在 GBS 患者的外周血中，而且在 EAN 大鼠中也观察到 P2X7R 的表达增加。用 BBG 抑制 P2X7R 可减轻 EAN 大鼠的神经症状，减少脱髓鞘，减少周围神经的炎性细胞浸润，并改善神经传导。BBG 还限制了促炎分子的产生，下调了 P2X7R 和 NLRP3 的表达，并抑制了 Th1 和 Th17 细胞的分化，从而保护 EAN 大鼠免受其影响。这些作用共同有助于改变 EAN 大鼠的炎症环境并改善其预后。
抑制 P2X7R 通过调节 CD4 T 细胞分化和 NLRP3 炎性小体激活来缓解 EAN 表现。这一发现突显了 P2X7R 作为抗炎治疗靶点的潜力，为 GBS 的研究和管理提供了新的思路。
