Kamat Pradip K, Khan Mohammad Badruzzaman, Siddiqui Shahneela, Hattaway Tyler Grace, Anas Affan, Rudic R Daniel, Baban Babak, Dhandapani Krishnan M, Hess David C
Departments of Neurology, Medical College of Georgia, Augusta University, Augusta, USA.
Departments of Neurology, Medical College of Georgia, Augusta University, Augusta, USA.
Exp Neurol. 2025 Feb;384:115067. doi: 10.1016/j.expneurol.2024.115067. Epub 2024 Nov 17.
Ischemic stroke leads to disability and death worldwide and evidence suggests that stroke severity is affected by the time dimension of the stroke. Rev-Erbα regulates the core circadian clock through repression of the positive clock element Bmal1. However, it remains unclear if a Rev-Erbα agonist (SR9009) alleviates stroke pathology in mice. We found that stroke reduces the level of Rev-Erbα and elevates neuroinflammation and stroke severity at zeitgeber time (ZT) ZT06. Therefore, we hypothesized that SR9009 treatment may reduce neuroinflammation and stroke severity in a mouse suture occlusion model. At 12 to 14 weeks, C57BL/6 J (Wild Type, n = 5-10 mice/group) mice were randomly assigned to undergo MCAO stroke for 60 min at either zeitgeber time ZT06 (MCAO-ZT06-sleep phase) or ZT18 (MCAO-ZT18-awake phase). Stroked mice were treated with SR9009 (100 mg/kg) or vehicle at 1 h and 24 h after MCAO. After forty-eight hours of stroke, TTC staining, Western blot, and qRT-PCR were performed. We found that SR9009 treatment alleviates neuroinflammation and infarct volume by Rev-Erb remodeling in ZT06 stroke mice but not in ZT18 stroke mice. Additionally, monocytic and neutrophilic NLRP3 as well as brain NLRP3 levels were reduced by SR9009 treatment in ZT06 stroke though no effects were observed at ZT18 stroke. SR9009 also reduced TNFα expression and increased IL-10 expression in blood and brain in ZT06 stroke mice and no differences were observed at ZT18. There were no significant effects of SR9009 on neurological deficit score and sensorimotor function at ZT06 or ZT18 at 48 h. Our study demonstrates that SR9009 treatment reduces stroke volume, circulating immune response, circadian expression, and that the protection was circadian- and treatment time-dependent.
缺血性中风在全球范围内导致残疾和死亡，有证据表明中风严重程度受中风时间维度的影响。Rev-Erbα通过抑制正向生物钟元件Bmal1来调节核心生物钟。然而，Rev-Erbα激动剂（SR9009）是否能减轻小鼠的中风病理仍不清楚。我们发现中风会降低Rev-Erbα水平，并在授时因子时间（ZT）ZT06时加剧神经炎症和中风严重程度。因此，我们假设SR9009治疗可能会减轻小鼠缝线闭塞模型中的神经炎症和中风严重程度。在12至14周龄时，将C57BL/6 J（野生型，每组n = 5 - 10只小鼠）小鼠随机分配，在授时因子时间ZT06（MCAO-ZT06-睡眠阶段）或ZT18（MCAO-ZT18-清醒阶段）接受60分钟的大脑中动脉闭塞（MCAO）中风。中风小鼠在MCAO后1小时和24小时接受SR9009（100 mg/kg）或赋形剂治疗。中风48小时后，进行TTC染色、蛋白质印迹和qRT-PCR。我们发现，SR9009治疗通过Rev-Erb重塑减轻了ZT06中风小鼠的神经炎症和梗死体积，但对ZT18中风小鼠无效。此外，SR9009治疗在ZT06中风时降低了单核细胞和中性粒细胞的NLRP3以及大脑中的NLRP3水平，而在ZT18中风时未观察到影响。SR9009还降低了ZT06中风小鼠血液和大脑中的TNFα表达，并增加了IL-10表达，在ZT18时未观察到差异。在48小时时，SR9009对ZT06或ZT18时的神经功能缺损评分和感觉运动功能没有显著影响。我们的研究表明，SR9009治疗可减少中风体积、循环免疫反应、昼夜节律表达，且这种保护作用具有昼夜节律和治疗时间依赖性。
