Amyotrophic lateral sclerosis (ALS) is a fatal and incurable disease involving motor neuron (MN) degeneration and is characterized by ongoing myasthenia and amyotrophia in adults. Most ALS patients die of respiratory muscle paralysis after an average of 3–5 years. Defective autophagy in MNs is considered an important trigger of ALS pathogenesis. Roflupram (ROF) was demonstrated to activate autophagy in microglial cells and exert protective effects against Parkinson's disease (PD) and Alzheimer's disease (AD). Therefore, our research aimed to investigate the efficacy and mechanism of ROF in treating ALS both in vivo and in vitro. We found that ROF could delay disease onset and prolong the survival of hSOD1-G93A transgenic mice. Moreover, ROF protected MNs in the anterior horn of the spinal cord, activated the AMPK/ULK1 signaling pathway, increased autophagic flow, and reduced SOD1 aggregation. In an NSC34 cell line stably transfected with hSOD1-G93A, ROF protected against cellular damage caused by hSOD1-G93A. Moreover, we have demonstrated that ROF inhibited gliosis in ALS model mice. Collectively, our study suggested that ROF is neuroprotective in ALS models and the AMPK/ULK1 signaling pathway is a potential therapeutic target in ALS, which increases autophagic flow and reduces SOD1 aggregation.

肌萎缩侧索硬化症 (ALS) 是一种致命且无法治愈的疾病，涉及运动神经元 (MN) 变性，其特征是成人持续出现肌无力和肌萎缩。大多数 ALS 患者平均在 3-5 年后死于呼吸肌麻痹。 MN 中的自噬缺陷被认为是 ALS 发病机制的重要触发因素。 Roflupram (ROF) 被证明可以激活小胶质细胞的自噬，并对帕金森病 (PD) 和阿尔茨海默病 (AD) 发挥保护作用。因此，我们的研究旨在探讨ROF在体内和体外治疗ALS的功效和机制。我们发现ROF可以延缓疾病发作并延长hSOD1-G93A转基因小鼠的存活时间。此外，ROF 保护脊髓前角的 MN，激活 AMPK/ULK1 信号通路，增加自噬流，并减少 SOD1 聚集。在用hSOD1-G93A稳定转染的 NSC34 细胞系中，ROF 可以防止hSOD1-G93A引起的细胞损伤。此外，我们已经证明 ROF 抑制ALS 模型小鼠的神经胶质增生。总的来说，我们的研究表明 ROF 在 ALS 模型中具有神经保护作用，而 AMPK/ULK1 信号通路是 ALS 的潜在治疗靶点，可增加自噬流并减少 SOD1 聚集。