Zhang Xiaoyan, Zhang Wei, Wang Yanyun, Zhang Yun, Zhang Dunke, Qin Guangcheng, Zhou Jiying, Chen Lixue
Department of Neurology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
Laboratory Research Center, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
Front Mol Neurosci. 2024 May 22;17:1387481. doi: 10.3389/fnmol.2024.1387481. eCollection 2024.
Central sensitization is one of the pivotal pathological mechanisms in chronic migraine (CM). Silent information regulator 1 (SIRT1) was shown to be involved in CM, but its specific mechanism is unclear. Reactive oxygen species (ROS) are increasingly regarded as important signaling molecules in several models of pain. However, studies about the role of ROS in the central sensitization of CM model are rare. We thus explored the specific process of SIRT1 involvement in the central sensitization of CM, focusing on the ROS pathway.
Inflammatory soup was repeatedly administered to male Sprague-Dawley rats to establish a CM model. The SIRT1 expression level in trigeminal nucleus caudalis (TNC) tissues was assessed by qRT-PCR and Western blotting analysis. The levels of ROS were detected by a Tissue Reactive Oxygen Detection Kit, DHE staining, and the fluorescence signal intensity of 8-OHdG. A ROS scavenger (tempol), a SIRT1 activator (SRT1720), a SIRT1 inhibitor (EX527), and a mitochondrial fission inhibitor (Mdivi-1) were used to investigate the specific molecular mechanisms involved. NMDAR2B, CGRP, ERK, and mitochondrial fission-related protein were evaluated by Western blotting, and the CGRP level in frozen sections of the TNC was detected via immunofluorescence staining.
After repeated inflammatory soup infusion and successful establishment of the CM rat model, SIRT1 expression was found to be significantly reduced, accompanied by elevated ROS levels. Treatment with Tempol, SRT1720, or Mdivi-1 alleviated allodynia and reduced the increase in NMDAR2B phosphorylation and CGRP and ERK phosphorylation in the CM rat. In contrast, EX527 had the opposite effect in CM rat. SRT1720 and EX527 decreased and increased ROS levels, respectively, in CM rats, and tempol reversed the aggravating effect of EX527 in CM rats. Furthermore, the regulatory effect of SIRT1 on ROS may include the involvement of the mitochondrial fission protein DRP1.
The results indicate the importance of SIRT1 in CM may be due to its role in regulating the production of ROS, which are involved in modulating central sensitization in CM. These findings could lead to new ideas for CM treatment with the use of SIRT1 agonists and antioxidants.
中枢敏化是慢性偏头痛（CM）的关键病理机制之一。沉默信息调节因子1（SIRT1）被证明与CM有关，但其具体机制尚不清楚。活性氧（ROS）在多种疼痛模型中越来越被视为重要的信号分子。然而，关于ROS在CM模型中枢敏化中的作用的研究很少。因此，我们探讨了SIRT1参与CM中枢敏化的具体过程，重点关注ROS途径。
对雄性Sprague-Dawley大鼠反复给予炎性介质以建立CM模型。通过qRT-PCR和蛋白质免疫印迹分析评估三叉神经脊束核（TNC）组织中SIRT1的表达水平。使用组织活性氧检测试剂盒、DHE染色和8-OHdG的荧光信号强度检测ROS水平。使用ROS清除剂（tempol）、SIRT1激活剂（SRT1720）、SIRT1抑制剂（EX527）和线粒体分裂抑制剂（Mdivi-1）来研究其中涉及的具体分子机制。通过蛋白质免疫印迹评估NMDAR2B、降钙素基因相关肽（CGRP）、细胞外信号调节激酶（ERK）和线粒体分裂相关蛋白，并通过免疫荧光染色检测TNC冰冻切片中的CGRP水平。
在反复注入炎性介质并成功建立CM大鼠模型后，发现SIRT1表达显著降低，同时ROS水平升高。用tempol、SRT1720或Mdivi-1治疗可减轻CM大鼠的痛觉过敏，并降低NMDAR2B磷酸化、CGRP和ERK磷酸化的增加。相反，EX527在CM大鼠中具有相反的作用。SRT1720和EX527分别降低和升高了CM大鼠的ROS水平，tempol逆转了EX527对CM大鼠的加重作用。此外，SIRT1对ROS的调节作用可能包括线粒体分裂蛋白动力相关蛋白1（DRP1）的参与。
结果表明SIRT1在CM中的重要性可能归因于其在调节ROS产生中的作用，而ROS参与调节CM中的中枢敏化。这些发现可能为使用SIRT1激动剂和抗氧化剂治疗CM带来新的思路。
