这项研究调查了唑尼沙胺治疗对实验性酒精成瘾(AA)模型中小脑组织的影响及其潜在的作用机制,特别是关于凋亡蛋白酶活化因子-1(APAF-1)和肿瘤坏死因子-α(TNF-α)的表达。30只大鼠分为三组:假,乙醇(EtOH),和EtOH+唑尼沙胺。通过每8小时口服施用6cc的EtOH诱导AA,持续4天。在给药EtOH之前,每天一次给予大鼠唑尼沙胺(100mg/kg)。使用开场迷宫评估电机缺陷。从血液样品中测量血清TNF-α水平。对小脑切片进行组织学检查,并对APAF-1和TNF-α进行免疫染色。使用Cytoscape构建蛋白质相互作用网络,和功能注释用ShinyGO(0.80版)软件进行。与假手术组相比,EtOH组的行进面积显著减少(p=0.0005)。EtOH+唑尼沙胺组大鼠覆盖面积较大,与EtOH组相比,唑尼沙胺治疗可显着改善运动能力(p=0.0463)。与假手术组相比,EtOH组的血清TNF-α水平显着升高(p<0.0001),而与EtOH组相比,EtOH唑尼沙胺组的血清TNF-α水平显着降低(p=0.0309)。在假手术组中观察到有规律的小脑组织学层,而EtOH诱导导致小脑组织完整性丧失,神经元变性,血管扩张和充血,髓鞘密度降低,和EtOH组的神经痛。唑尼沙胺治疗改善了这些病变,增强髓鞘形成和神经纤维形成。假手术组大鼠小脑层APAF-1和TNF-α呈阴性表达。由于EtOH的毒性,与假手术组相比,EtOH组的APAF-1和TNF-α表达上调(两者p<0.001)。与EtOH组相比,唑尼沙胺处理下调EtOH+唑尼沙胺组中的这些蛋白质表达(分别为p<0.001和p=0.0087)。APAF-1主要通过抗叶酸剂与AA相关,内肽酶,和白细胞介素-1通路,而TNF-α主要富集在感染和胆碱结合中,表明唑尼沙胺对免疫和炎症途径的影响。总之,唑尼沙胺治疗可显着减轻AA模型中乙醇诱导的小脑损伤和炎症。唑尼沙胺改善运动功能,降低血清TNF-α水平,以及APAF-1和TNF-α在小脑组织中的表达。这些发现表明唑尼沙胺通过调节免疫和炎症途径发挥其保护作用,从而保持小脑的完整性和功能。
This study investigated the effects of
zonisamide treatment on cerebellar tissues in an experimental alcohol addiction (AA) model and its potential mechanisms of action, particularly regarding apoptotic protease activating factor-1 (APAF-1) and tumor necrosis factor-alpha (TNF-α) expression. Thirty rats were divided into three groups: sham, ethanol (EtOH), and EtOH +
zonisamide. AA was induced by administering 6 cc of EtOH orally every 8 h for 4 days.
Zonisamide (100 mg/kg) was given to rats once daily before EtOH administration. Motor defects were evaluated using an open field maze. Serum TNF-α levels were measured from blood samples. Cerebellar sections were processed for histological examination and immunostained for APAF-1 and TNF-α. Protein interaction networks were constructed using Cytoscape, and functional annotations were performed with ShinyGO (version 0.80) software. The traveled area in the EtOH group was significantly reduced compared to the sham group (p = 0.0005). Rats in the EtOH +
zonisamide group covered a larger area, with
zonisamide treatment significantly improving locomotor ability compared to the EtOH group (p = 0.0463). Serum TNF-α levels were significantly elevated in the EtOH group compared to the sham group (p < 0.0001) and were significantly decreased in the EtOH + zonisamide group compared to the EtOH group (p = 0.0309). Regular cerebellar histological layers were observed in the sham group, while EtOH induction caused loss of cerebellar tissue integrity, neuronal degeneration, vascular dilatation and congestion, reduced myelin density, and neuropils in the EtOH group. Zonisamide treatment improved these pathologies, enhancing myelination and neuropil formation. Negative APAF-1 and TNF-α expressions were observed across cerebellar layers in the sham group. Due to EtOH toxicity, APAF-1 and TNF-α expression were upregulated in the EtOH group compared to the sham group (p < 0.001 for both).
Zonisamide treatment downregulated these protein expressions in the EtOH +
zonisamide group compared to the EtOH group (p < 0.001 and p = 0.0087, respectively). APAF-1 was primarily associated with AA through antifolate resistance, endopeptidases, and the interleukin-1 pathway, while TNF-α was predominantly enriched in infections and choline-binding, indicating zonisamide\'s impact on immune and inflammatory pathways. In conclusion, zonisamide treatment significantly mitigated ethanol-induced cerebellar damage and inflammation in an AA model. Zonisamide improved locomotor function and reduced serum TNF-α levels, as well as APAF-1 and TNF-α expression in cerebellar tissues. These findings suggest that zonisamide exerts its protective effects by modulating immune and inflammatory pathways, thereby preserving cerebellar integrity and function.