Li Jie, Wang Qian, Gao Yangzheng, Ma Wanjie, Sun Zexian, Yu Yonghao, Li Yize, Li Qing, Wang Chunyan
Department of Anesthesiology, Tianjin Medical University General Hospital, Tianjin 300052, PR China; Tianjin Research Institute of Anesthesiology, Tianjin 300052, PR China.
Department of Anesthesiology, Tianjin Medical University General Hospital, Tianjin 300052, PR China; Tianjin Research Institute of Anesthesiology, Tianjin 300052, PR China.
Neurosci Lett. 2023 Mar 28;801:137131. doi: 10.1016/j.neulet.2023.137131. Epub 2023 Feb 15.
Recent research shows a correlation between altered sphingolipid metabolism and nociceptive processing. Activation of the sphingosine-1-phosphate receptor 1 subtype (S1PR1) by its ligand, sphingosine-1-phosphate (S1P), causes neuropathic pain. However, its role in remifentanil-induced hyperalgesia (RIH) has not been investigated. The purpose of this research was to establish if the SphK/S1P/S1PR1 axis mediated remifentanil-induced hyperalgesia and identify its potential targets. This study examined the protein expression of ceramide, sphingosine kinases (SphK), S1P, and S1PR1 in the spinal cord of rats treated with remifentanil (1.0 μg/kg/min for 60 min). Prior to receiving remifentanil, rats were injected with SK-1 (a SphK inhibitor); LT1002 (a S1P monoclonal antibody); CYM-5442, FTY720, and TASP0277308（the S1PR1 antagonists); CYM-5478 (a S1PR2 agonist); CAY10444 (a S1PR3 antagonist); Ac-YVAD-CMK (a caspase-1 antagonist); MCC950 (the NOD-like receptor protein 3 (NLRP3) inflammasome antagonist); and N-tert-Butyl-α-phenylnitrone (PBN, a reactive oxygen species (ROS) scavenger). Mechanical and thermal hyperalgesia were evaluated at baseline (24 h prior to remifentanil infusion) and 2, 6, 12, and 24 h following remifentanil administration. The expression of the NLRP3-related protein (NLRP3, caspase-1), pro-inflammatory cytokines (interleukin-1β(IL-1β), IL-18), and ROS was found in the spinal dorsal horns. In the meantime, immunofluorescence was used to ascertain if S1PR1 co-localizes with astrocytes. Remifentanil infusion induced considerable hyperalgesia in addition to increased ceramide, SphK, S1P, and S1PR1, NLRP3-related protein (NLRP3, Caspase-1, IL-1β, IL-18) and ROS expression, and S1PR1 localized astrocytes. By blocking the SphK/S1P/S1PR1 axis, remifentanil-induced hyperalgesia was reduced, as was the expression of NLRP3, caspase-1, pro-inflammatory cytokines (IL-1β, IL-18) and ROS in the spinal cord. In addition, we observed that suppressing NLRP3 or ROS signal attenuated the mechanical and thermal hyperalgesia induced by remifentanil. Our findings indicate that the SphK/SIP/S1PR1 axis regulates the expression of NLRP3, Caspase-1, IL-1β, IL-18 and ROS in the spinal dorsal horn to mediate remifentanil-induced hyperalgesia. These findings may contribute to pain and SphK/S1P/S1PR1 axis research positively, and inform the future study of this commonly used analgesic.
近期研究表明，鞘脂代谢改变与伤害性感受处理之间存在关联。鞘氨醇-1-磷酸（S1P）作为其配体激活1型鞘氨醇-1-磷酸受体（S1PR1）会引发神经性疼痛。然而，其在瑞芬太尼诱发痛觉过敏（RIH）中的作用尚未得到研究。本研究的目的是确定鞘氨醇激酶（SphK）/S1P/S1PR1轴是否介导瑞芬太尼诱发的痛觉过敏，并确定其潜在靶点。本研究检测了接受瑞芬太尼（1.0μg/kg/分钟，持续60分钟）治疗的大鼠脊髓中神经酰胺、鞘氨醇激酶（SphK）、S1P和S1PR1的蛋白表达。在接受瑞芬太尼之前，给大鼠注射SK-1（一种SphK抑制剂）；LT1002（一种S1P单克隆抗体）；CYM-5442、FTY720和TASP0277308（S1PR1拮抗剂）；CYM-5478（一种S1PR2激动剂）；CAY10444（一种S1PR3拮抗剂）；Ac-YVAD-CMK（一种半胱天冬酶-1拮抗剂）；MCC950（NOD样受体蛋白3（NLRP3）炎性小体拮抗剂）；以及N-叔丁基-α-苯基硝酮（PBN，一种活性氧（ROS）清除剂）。在基线（瑞芬太尼输注前24小时）以及瑞芬太尼给药后2、6、12和24小时评估机械性和热痛觉过敏。在脊髓背角发现了NLRP3相关蛋白（NLRP3、半胱天冬酶-1）、促炎细胞因子（白细胞介素-1β（IL-1β）、IL-18）和ROS的表达。同时，使用免疫荧光法确定S1PR1是否与星形胶质细胞共定位。输注瑞芬太尼除了会导致神经酰胺、SphK、S1P和S1PR1、NLRP3相关蛋白（NLRP3、半胱天冬酶-1、IL-1β、IL-18）以及ROS表达增加外，还会诱发明显的痛觉过敏，并且S1PR1定位于星形胶质细胞。通过阻断SphK/S1P/S1PR1轴，瑞芬太尼诱发的痛觉过敏以及脊髓中NLRP3、半胱天冬酶-1、促炎细胞因子（IL-1β、IL-18）和ROS的表达均降低。此外，我们观察到抑制NLRP3或ROS信号可减轻瑞芬太尼诱发的机械性和热痛觉过敏。我们的研究结果表明，SphK/SIP/S1PR1轴调节脊髓背角中NLRP3、半胱天冬酶-1、IL-1β、IL-18和ROS的表达，以介导瑞芬太尼诱发的痛觉过敏。这些发现可能会对疼痛和SphK/S1P/S1PR1轴的研究产生积极贡献，并为这种常用镇痛药的未来研究提供参考。
