Alveolar macrophages (AMs) are the primary targets of silicosis. Blockade of autophagy may aggravate the apoptosis of AMs. Trehalose (Tre), a transcription factor EB (TFEB) activator, may impact the autophagy-lysosomal system in AMs during silicosis. However, the mechanism by which Tre acts upon AMs in silicosis is unknown. We collected AMs from twenty male workers exposed to silica and divided them into observer and silicosis patient groups. AMs from the two groups were then exposed to Tre. Western blot was used to measure the expression of autophagy-associated proteins. Lysosomal-associated membrane protein 1 (LAMP1) expression was observed using immunofluorescence and western blot. Apoptosis of the AMs was detected by TUNEL assay and western blot. Tre induced localization of TFEB to the nucleus in the AMs of both groups. After Tre exposure, LAMP1 levels increased and LC3 levels decreased in the AMs of both groups, suggesting that Tre may increase the function of the autophagy-lysosomal system. The LC3-II/I ratio in the Tre-exposed AMs was lower than in the AMs not exposed to Tre. The LC3-II/I ratio in AMs subjected to Tre plus Bafilomycin (Baf) was higher than the ratio in cells exposed to Tre or Baf individually. Additionally, p62 levels decreased after Tre stimulation in the AMs of both groups. This indicates that Tre may accelerate the process of autophagic degradation. We also found decreased levels of cleaved caspase-3 after Tre treatment in the AMs of both groups. However, p-mTOR (Ser2448) and p-mTOR (Ser2481) levels did not change significantly after Tre treatment, suggesting that the mTOR signaling pathway was not affected by Tre treatment. Our findings suggest that the restoration of autophagy-lysosomal function by Tre may be a potential protective strategy against silicosis.

中文翻译：

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海藻糖通过保护人矽肺期间肺泡巨噬细胞的自噬-溶酶体系统来减轻细胞凋亡

肺泡巨噬细胞（AM）是矽肺的主要靶标。阻断自噬可能会加剧 AMs 的凋亡。海藻糖 (Tre) 是一种转录因子 EB (TFEB) 激活剂，可能会影响矽肺期间 AM 的自噬-溶酶体系统。然而，Tre 在矽肺中作用于 AM 的机制尚不清楚。我们收集了 20 名接触二氧化硅的男性工人的 AM，并将他们分为观察组和矽肺患者组。然后将两组的 AM 暴露于 Tre。蛋白质印迹用于测量自噬相关蛋白的表达。使用免疫荧光和蛋白质印迹观察溶酶体相关膜蛋白 1 (LAMP1) 的表达。通过TUNEL法和蛋白质印迹法检测AM的凋亡。 Tre 诱导 TFEB 定位至两组 AM 的细胞核。 Tre暴露后，两组AM中LAMP1水平升高，LC3水平降低，表明Tre可能增强自噬-溶酶体系统的功能。暴露于 Tre 的 AM 中的 LC3-II/I 比值低于未暴露于 Tre 的 AM 中。接受 Tre 加巴弗洛霉素 (Baf) 的 AM 中的 LC3-II/I 比率高于单独暴露于 Tre 或 Baf 的细胞中的比率。此外，两组 AM 中的 Tre 刺激后 p62 水平均下降。这表明Tre可能加速自噬降解的过程。我们还发现 Tre 治疗后两组 AM 中裂解的 caspase-3 水平均下降。然而，Tre治疗后p-mTOR（Ser2448）和p-mTOR（Ser2481）水平没有显着变化，表明mTOR信号通路不受Tre治疗的影响。我们的研究结果表明，Tre 恢复自噬溶酶体功能可能是预防矽肺的潜在保护策略。