背景:临床上,缺血再灌注损伤是中风损伤的主要原因。本研究旨在评估芬戈莫德抑制缺血性脑损伤引起的炎症反应的有效性,并探讨其药理机制。
方法:总共,将75只雄性Sprague-Dawley大鼠随机平均分为五个不同的组:假,大脑中动脉闭塞/再灌注(MCAO/R)手术,芬戈莫德低剂量(F-L),芬戈莫德中等剂量(F-M),和芬戈莫德高剂量(F-H)。神经行为测试,氯化2,3,5-三苯基四唑染色,并采用脑组织干湿法评估神经功能缺损,脑梗死大小,和大脑含水量。酶联免疫吸附试验用于定量促炎细胞因子白细胞介素(IL)-1β,IL-6和肿瘤坏死因子-α(TNF-α)蛋白水平。进行蛋白质印迹和免疫组织化学染色以评估高迁移率组1(HMGB1),toll样受体4(TLR4),核因子κBp65(NF-κBp65)水平。
结果:F-L中的大鼠,F-M,F-H组Longa评分较低,梗死体积减少,脑水肿较MCAO/R组减轻。此外,F-L,F-M,F-H组血清IL-1β水平较低,IL-6、TNF-α高于MCAO/R组。此外,F-L,F-M,和F-H处理导致MCAO/R大鼠海马HMGB1、TLR4和NF-κBp65蛋白表达水平降低。
结论:发现芬戈莫以剂量依赖性方式减轻缺血性脑损伤。此外,还发现它通过HMGB1/TLR4/NF‑κB信号通路减轻缺血性脑损伤后的炎症。
BACKGROUND: Clinically, ischemic reperfusion injury is the main cause of stroke injury. This study aimed to assess the effectiveness of fingolimod in suppressing inflammation caused by ischemic brain injury and explore its pharmacological mechanisms.
METHODS: In total, 75 male Sprague-Dawley rats were randomly and equally assigned to five distinct groups: sham, middle cerebral artery occlusion/reperfusion (MCAO/R) surgery, fingolimod low-dose (F-L), fingolimod medium-dose (F-M), and fingolimod high-dose (F-H). Neurobehavioral tests, 2,3,5-triphenyltetrazolium chloride staining, and the brain tissue drying-wet method were conducted to evaluate neurological impairment, cerebral infarction size, and brain water content. Enzyme-linked immunosorbent assay was employed to quantify pro-inflammatory cytokines interleukin (IL)-1β, IL-6, and tumor necrosis factor-alpha (TNF-α) protein levels. Western blotting and immunohistochemical staining were performed to assess high mobility group box 1 (HMGB1), toll-like receptor 4 (TLR4), and nuclear factor kappa-B p65 (NF-κBp65) levels.
RESULTS: Rats in the F-L, F-M, and F-H groups exhibited lower Longa scores, reduced infarction volumes, and decreased brain edema than those in the MCAO/R group. Additionally, the F-L, F-M, and F-H groups exhibited lower serum levels of IL-1β, IL-6, and TNF-α than those of the MCAO/R group. Additionally, F-L, F-M, and F-H treatments resulted in decreased HMGB1, TLR4, and NF-κBp65 protein expression levels in the hippocampus of MCAO/R rats.
CONCLUSIONS: Fingolimod was found to reduce ischemic brain injury in a dose-dependent manner. Moreover, it was also found to alleviate inflammation following ischemic brain injury via the HMGB1/TLR4/NF‑κB signaling pathway.