Martínez-Torres Ari Misael, Ramírez-Celis Crisalde, Morán Julio
División de Neurociencias, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Ciudad de México, México.
CNS Neurosci Ther. 2025 Apr;31(4):e70385. doi: 10.1111/cns.70385.
Intracerebral hemorrhage (ICH) is a leading cause of death and disability worldwide. Following the initial mechanical injury caused by hematoma expansion, a secondary injury occurs, characterized by the production of reactive oxygen species (ROS) generated by NOX-2 and neuroinflammation, which is exacerbated by the upregulation of the NLRP3 inflammasome. These conditions collectively aggravate brain damage. The endocannabinoid system (ECS), through the activation of the cannabinoid receptors, has demonstrated neuroprotective properties in various models of brain injury. However, the role of the ECS during ICH remains poorly understood, particularly regarding the action of the CB1 receptor in the activation of NOX-2 and the inflammasome. The present study investigates the neuroprotective effects of the cannabinoid receptor agonist WIN55,212-2 in an ICH animal model, specifically examining the roles of NLRP3 and NOX-2.
Male C57BL/6 mice were subjected to ICH through an intracerebral injection of collagenase, followed by intraperitoneal administration of WIN55,212-2 and/or MCC950, a selective NLRP3 inhibitor. Various outcome measures were employed, including assessments of motor activity, hematoma volume, brain water content, and blood-brain barrier (BBB) permeability, which was evaluated using Evans blue assay. Additionally, the activity of NOX and the protein levels of crucial markers such as CB1, gp91phox, NLRP3, AQP4, and caspase-1 were measured via western blot analysis.
The findings demonstrate that ICH induced a significant brain lesion characterized by hematoma formation, edema, BBB disruption, and subsequent motor impairments in the affected mice. Notably, these detrimental effects were markedly reduced in animals treated with WIN55,212-2. The study also revealed an activation of both NOX-2 and NLRP3 in response to ICH, which was reduced by cannabinoid receptor activation. Furthermore, the pharmacological inhibition of NLRP3 using MCC950 also led to a reduction in hematoma size, edema, and motor impairment secondary to ICH.
These results support a neuroprotective role of the cannabinoid receptor activation during ICH and suggest the involvement of NOX-2 and NLRP3.
脑出血（ICH）是全球范围内导致死亡和残疾的主要原因。在血肿扩大引起的初始机械性损伤之后，会发生继发性损伤，其特征是由NOX-2产生的活性氧（ROS）和神经炎症，而NLRP3炎性小体的上调会加剧这种炎症。这些情况共同加重脑损伤。内源性大麻素系统（ECS）通过激活大麻素受体，在各种脑损伤模型中已显示出神经保护特性。然而，ECS在脑出血期间的作用仍知之甚少，特别是关于CB1受体在激活NOX-2和炎性小体中的作用。本研究调查了大麻素受体激动剂WIN55,212-2在脑出血动物模型中的神经保护作用，特别研究了NLRP3和NOX-2的作用。
雄性C57BL/6小鼠通过脑内注射胶原酶诱导脑出血，随后腹腔注射WIN55,212-2和/或MCC950（一种选择性NLRP3抑制剂）。采用了各种结局指标，包括运动活动评估、血肿体积、脑含水量和血脑屏障（BBB）通透性评估（使用伊文思蓝试验进行评估）。此外，通过蛋白质免疫印迹分析测量了NOX的活性以及CB1、gp91phox、NLRP3、AQP4和caspase-1等关键标志物的蛋白质水平。
研究结果表明，脑出血在受影响的小鼠中诱发了以血肿形成、水肿、血脑屏障破坏和随后的运动障碍为特征的显著脑损伤。值得注意的是，在用WIN55,212-2治疗的动物中，这些有害影响明显减轻。该研究还揭示，脑出血会激活NOX-2和NLRP3，而大麻素受体激活会使其减少。此外，使用MCC950对NLRP3进行药理抑制也导致脑出血继发的血肿大小、水肿和运动障碍有所减轻。
这些结果支持大麻素受体激活在脑出血期间具有神经保护作用，并提示NOX-2和NLRP3参与其中。
