Department of Surgical and Interventional Sciences, McGill University, Montreal, QC H3T 1E2, Canada.
Orthopaedic Research Laboratory, Lady Davis Institute for Medical Research, Montreal, QC H3T 1E2, Canada.
Biomolecules. 2024 Oct 16;14(10):1312. doi: 10.3390/biom14101312.
Intervertebral disc degeneration and pain are associated with the nucleotide-binding domain, leucine-rich repeat, and pyrin domain-containing 3 (NLRP3) inflammasome activation and the processing of interleukin-1 beta (IL-1β). Activation of thehm inflammasome is triggered by Toll-like receptor stimulation and requires the cofactor receptor cluster of differentiation 14 (CD14). Short Link N (sLN), a peptide derived from link protein, has been shown to modulate inflammation and pain in discs in vitro and in vivo; however, the underlying mechanisms remain elusive. This study aims to assess whether sLN modulates IL-1β and inflammasome activity through interaction with CD14. Disc cells treated with lipopolysaccharides (LPS) with or without sLN were used to assess changes in Caspase-1, IL-1β, and phosphorylated nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB). Peptide docking of sLN to CD14 and immunoprecipitation were performed to determine their interaction. The results indicated that sLN inhibited LPS-induced NFκB and Caspase-1 activation, reducing IL-1β maturation and secretion in disc cells. A significant decrease in inflammasome markers was observed with sLN treatment. Immunoprecipitation studies revealed a direct interaction between sLN and the LPS-binding pocket of CD14. Our results suggest that sLN could be a potential therapeutic agent for discogenic pain by mitigating IL-1β and inflammasome activity within discs.
椎间盘退变和疼痛与核苷酸结合域、富含亮氨酸重复和吡喃结构域蛋白 3（NLRP3）炎症小体激活以及白细胞介素 1β（IL-1β）的加工有关。炎症小体的激活是由 Toll 样受体刺激触发的，需要共受体分化簇 14（CD14）。短链 N（sLN）是一种来源于连接蛋白的肽，已被证明可以在体外和体内调节椎间盘的炎症和疼痛；然而，其潜在机制仍不清楚。本研究旨在评估 sLN 是否通过与 CD14 的相互作用来调节 IL-1β 和炎症小体活性。用脂多糖（LPS）处理或不处理 sLN 的椎间盘细胞用于评估 Caspase-1、IL-1β 和磷酸化核因子 kappa-轻链增强子活化 B 细胞（NFκB）的变化。进行 sLN 与 CD14 的肽对接和免疫沉淀以确定它们的相互作用。结果表明，sLN 抑制 LPS 诱导的 NFκB 和 Caspase-1 激活，减少椎间盘细胞中 IL-1β 的成熟和分泌。sLN 处理后炎症小体标志物明显减少。免疫沉淀研究显示 sLN 与 CD14 的 LPS 结合口袋之间存在直接相互作用。我们的结果表明，sLN 可能通过减轻椎间盘内的 IL-1β 和炎症小体活性成为治疗椎间盘源性疼痛的潜在治疗剂。
