目的:肌萎缩侧索硬化症(ALS)是一种严重的神经退行性疾病,其特征是上,下运动神经元进行性死亡,导致全身肌肉萎缩,瘫痪甚至死亡。线粒体损伤和神经炎症在ALS的发病机制中起关键作用。在本研究中,A-1是具有AMP激活的蛋白激酶(AMPK)和沉默信息调节因子1(SIRT1)激活的牛肝素元衍生物,被调查。
方法:从第13周开始口服给SOD1G93A转基因小鼠施用33.3mg/kg的A-1,持续6周的治疗期。在麻醉结束前评估运动能力。肌肉萎缩和纤维化,运动神经元,星形胶质细胞,通过H&E评估脊髓中的小胶质细胞,Masson,小天狼星红,Nissl,和免疫组织化学染色。用蛋白质组学分析检测蛋白质表达,西方印迹,和ELISA。使用测定试剂盒测量线粒体三磷酸腺苷(ATP)和丙二醛(MDA)水平。
结果:在SOD1G93A小鼠中给予A-1增强了活动性,减少骨骼肌萎缩和纤维化,减轻脊髓运动神经元的损失,并减少神经胶质的激活。此外,A-1治疗改善线粒体功能,ATP水平升高和关键线粒体相关蛋白表达增加。A-1治疗组IL-1β水平下降,pIκBα/IκBα,和pNF-κB/NF-κB。
结论:A-1治疗减少了运动神经元的损失,改善腓肠肌萎缩,并通过AMPK/SIRT1/PGC-1α途径延迟ALS进展,促进线粒体生物发生。此外,AMPK/SIRT1/IL-1β/NF-κB通路通过减轻神经炎症而发挥神经保护作用。这些发现表明A-1是ALS的有希望的治疗方法。
OBJECTIVE: Amyotrophic lateral sclerosis (ALS) is a severe neurodegenerative disease characterized by progressive death of upper and lower motor neurons, leading to generalized muscle atrophy, paralysis, and even death. Mitochondrial damage and neuroinflammation play key roles in the pathogenesis of ALS. In the present study, the efficacy of A-1, a derivative of arctigenin with AMP-activated protein kinase (AMPK) and silent information regulator 1 (SIRT1) activation for ALS, was investigated.
METHODS: A-1 at 33.3 mg/kg was administrated in SOD1G93A transgenic mice orally from the 13th week for a 6-week treatment period. Motor ability was assessed before terminal anesthesia. Muscle atrophy and fibrosis, motor neurons, astrocytes, and microglia in the spinal cord were evaluated by H&E, Masson, Sirius Red, Nissl, and immunohistochemistry staining. Protein expression was detected with proteomics analysis, Western blotting, and ELISA. Mitochondrial adenosine triphosphate (ATP) and malondialdehyde (MDA) levels were measured using an assay kit.
RESULTS: A-1 administration in SOD1G93A mice enhanced mobility, decreased skeletal muscle atrophy and fibrosis, mitigated loss of spinal motor neurons, and reduced glial activation. Additionally, A-1 treatment improved mitochondrial function, evidenced by elevated ATP levels and increased expression of key mitochondrial-related proteins. The A-1 treatment group showed decreased levels of IL-1β, pIκBα/IκBα, and pNF-κB/NF-κB.
CONCLUSIONS: A-1 treatment reduced motor neuron loss, improved gastrocnemius atrophy, and delayed ALS progression through the AMPK/SIRT1/PGC-1α pathway, which promotes mitochondrial biogenesis. Furthermore, the AMPK/SIRT1/IL-1β/NF-κB pathway exerted neuroprotective effects by reducing neuroinflammation. These findings suggest A-1 as a promising therapeutic approach for ALS.