背景:已发现高频重复经颅磁刺激(HF-rTMS)可改善认知障碍。然而,HF-rTMS在慢性脑低灌注(CCH)中的作用仍然未知。
目的:研究HF-rTMS对CCH小鼠认知功能改善的影响及其可能机制。
方法:双侧颈动脉狭窄(BCAS)后每天进行HF-rTMS治疗并持续14天。将小鼠随机分为三组:假手术组,模型组,和HF-rTMS组。使用Y迷宫和新物体识别测试来评估认知功能。MAP-2的表达,突触,髓鞘碱性蛋白(MBP),通过免疫荧光染色和蛋白质印迹分析脑源性生长因子(BDNF),以评估神经元可塑性和白质髓鞘再生。Nissl染色和caspase-3,Bax的表达,用Bcl-2观察神经元凋亡。此外,通过荧光染色评估小胶质细胞和星形胶质细胞的活化。IL-1β的炎症水平,qPCR检测各组小鼠海马组织中IL-6和肿瘤坏死因子(TNF)-α的表达。
结果:通过行为测试,BCAS小鼠显示新对象偏好率降低,自发交替率降低,而HF-rTMS可显着改善海马学习和记忆障碍。此外,模型组小鼠MAP-2、突触、MBP,BDNF,而HF-rTMS治疗逆转了这些效应。不出所料,模型组激活的小胶质细胞和星形胶质细胞增多,但HF-rTMS治疗抑制了这些变化。HF-rTMS降低了BCAS诱导的神经元凋亡和促凋亡蛋白(Caspase-3和Bax)的表达,并增加了抗凋亡蛋白(Bcl-2)的表达。此外,HF-rTMS抑制炎性细胞因子(IL-1β,IL-6和TNF-α)。
结论:HF-rTMS通过增强神经元可塑性和抑制炎症反应减轻CCH小鼠的认知障碍,因此,作为血管性认知障碍的潜在方法。
BACKGROUND: High-frequency repetitive transcranial magnetic stimulation (HF-rTMS) has been found to ameliorate cognitive impairment. However, the effects of HF-rTMS remain unknown in chronic cerebral hypoperfusion (CCH).
OBJECTIVE: To investigate the effects of HF-rTMS on cognitive improvement and its potential mechanisms in CCH mice.
METHODS: Daily HF-rTMS therapy was delivered after bilateral carotid stenosis (BCAS) and continued for 14 days. The mice were randomly assigned to three groups: the sham group, the model group, and the HF-rTMS group. The Y maze and the new object recognition test were used to assess cognitive function. The expressions of MAP-2, synapsis, Myelin basic protein(MBP), and brain-derived growth factors (BDNF) were analyzed by immunofluorescence staining and western blot to evaluate neuronal plasticity and white matter myelin regeneration. Nissl staining and the expression of caspase-3, Bax, and Bcl-2 were used to observe neuronal apoptosis. In addition, the activation of microglia and astrocytes were evaluated by fluorescence staining. The inflammation levels of IL-1β, IL-6, and Tumor Necrosis Factor(TNF)-α were detected by qPCR in the hippocampus of mice in each group.
RESULTS: Via behavioral tests, the BCAS mice showed reduced a rate of new object preference and decreased a rate of spontaneous alternations, while HF-rTMS significantly improved hippocampal learning and memory deficits. In addition, the mice in the model group showed decreased levels of MAP-2, synapsis, MBP, and BDNF, while HF-rTMS treatment reversed these effects. As expected, activated microglia and astrocytes increased in the model group, but HF-rTMS treatment suppressed these changes. HF-rTMS decreased BCAS-induced neuronal apoptosis and the expression of pro-apoptotic protein (Caspase-3 and Bax) and increased the expression of anti-apoptotic protein (Bcl-2). In addition, HF-rTMS inhibited the expression of inflammatory cytokines (IL-1β, IL-6, and TNF-α).
CONCLUSIONS: HF-rTMS alleviates cognitive impairment in CCH mice by enhancing neuronal plasticity and inhibiting inflammation, thus serving as a potential method for vascular cognitive impairment.