神经性疼痛（NP）是各种神经系统疾病的主要症状。 NP 患者经常出现情绪障碍，特别是抑郁和焦虑，这会严重影响他们的正常生活。小胶质细胞与 NP 相关。过度的炎症反应，尤其是大量促炎细胞因子的分泌，最终导致神经炎症。小胶质细胞焦亡是一种新发现的炎症细胞死亡形式，与免疫反应和中枢神经系统炎症相关疾病相关。从NP和相关机制方面探讨A型肉毒杆菌毒素（BTX-A）对小胶质细胞焦亡的影响使用两种模型：体外脂多糖 (LPS) 刺激的小胶质细胞模型和使用 BTX-A 和 SPP1 敲低治疗的选择性神经损伤模型。使用蛋白质印迹、实时定量聚合酶链反应和免疫荧光评估细胞焦亡信号通路中的关键蛋白 NLRP3-GSDMD。使用酶联免疫吸附测定评估炎症因子[白细胞介素 (IL)-6、IL-1β 和肿瘤坏死因子 (TNF)-α]。我们还评估了小胶质细胞的增殖和凋亡。此外，我们通过使用热刺激评估延迟后爪缩回潜伏期来测量疼痛感觉。LPS 处理中 ACS 和 GSDMD-N 的表达水平以及 TNF-α、IL-6 和 IL-1β 的 mRNA 表达增强小胶质细胞。此外，LPS 处理的小胶质细胞中也诱导了 SPP1 表达。值得注意的是，BTX-A 抑制 LPS 处理的小胶质细胞中 SPP1 mRNA 和蛋白的表达。此外，SPP1或BTX-A的消耗会抑制LPS处理的小胶质细胞的细胞活力并诱导细胞凋亡，而与BTX-A的共同处理增强了LPS处理的小胶质细胞中SPP1短发夹（sh）RNA的作用。最后，SPP1耗竭或BTX-A治疗降低了GSDMD-N、NLPRP3和ASC的水平，并抑制炎症因子的产生。值得注意的是，BTX-A治疗和SPP1 shRNA增强小胶质细胞增殖和凋亡并抑制小胶质细胞死亡。它可以改善选择性神经痛大鼠的疼痛感知并抑制小胶质细胞激活。©作者2023。百事登出版集团有限公司出版。保留所有权利。

Neuropathic pain (NP) is the primary symptom of various neurological conditions. Patients with NP often experience mood disorders, particularly depression and anxiety, that can severely affect their normal lives. Microglial cells are associated with NP. Excessive inflammatory responses, especially the secretion of large amounts of pro-inflammatory cytokines, ultimately lead to neuroinflammation. Microglial pyroptosis is a newly discovered form of inflammatory cell death associated with immune responses and inflammation-related diseases of the central nervous system.To investigate the effects of botulinum toxin type A (BTX-A) on microglial pyroptosis in terms of NP and associated mechanisms.Two models, an in vitro lipopolysaccharide (LPS)-stimulated microglial cell model and a selective nerve injury model using BTX-A and SPP1 knockdown treatments, were used. Key proteins in the pyroptosis signaling pathway, NLRP3-GSDMD, were assessed using western blotting, real-time quantitative polymerase chain reaction, and immunofluorescence. Inflammatory factors [interleukin (IL)-6, IL-1β, and tumor necrosis factor (TNF)-α] were assessed using enzyme-linked immunosorbent assay. We also evaluated microglial cell proliferation and apoptosis. Furthermore, we measured pain sensation by assessing the delayed hind paw withdrawal latency using thermal stimulation.The expression levels of ACS and GSDMD-N and the mRNA expression of TNF-α, IL-6, and IL-1β were enhanced in LPS-treated microglia. Furthermore, SPP1 expression was also induced in LPS-treated microglia. Notably, BTX-A inhibited SPP1 mRNA and protein expression in the LPS-treated microglia. Additionally, depletion of SPP1 or BTX-A inhibited cell viability and induced apoptosis in LPS-treated microglia, whereas co-treatment with BTX-A enhanced the effect of SPP1 short hairpin (sh)RNA in LPS-treated microglia. Finally, SPP1 depletion or BTX-A treatment reduced the levels of GSDMD-N, NLPRP3, and ASC and suppressed the production of inflammatory factors.Notably, BTX-A therapy and SPP1 shRNA enhance microglial proliferation and apoptosis and inhibit microglial death. It improves pain perception and inhibits microglial activation in rats with selective nerve pain.©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.