Autophagy is a lysosomal degradation mechanism for elimination and recycling of damaged intracellular organelles and proteins. Recent studies have shown that autophagy could help reduce oxidative stress by removing oxidized proteins and damaged mitochondria. Autophagy deficiency is associated with the disruption of many intracellular biological processes. Using bioinformatics tools and fibroblast immunostaining technology, I tried to investigate whether oxidative stress is involved in mediating the effect of autophagy suppression on certain cell biological processes and signalling pathways. Many pharmaceutical components have different modes of action to suppress autophagy. In this study, I performed analysis on autophagy suppression induced by neutralizing lysosomal pH (NH₄Cl and bafilomycin A1). Bioinformatics analysis of GEO data, GSE60570 accession number, revealed that p38 signalling induction and DNA damage response are among the main disrupted signalling pathways in bafilomycin A1-treated RPE-1 cells. Likewise, fibroblast immunostaining showed that autophagy deficiency established by ammonium chloride (NH₄Cl) has significantly increased P38 signalling, DNA damage marker (H2A.X), and oxidative stress marker (dityrosine). I therefore investigated the role of oxidative stress and whether antioxidants treatment could reverse autophagy suppression effects on p38 signalling and DNA damage response. Importantly, antioxidant treatment clearly restored P38 signalling and H2A.X levels in autophagy-suppressed fibroblast cells. Indicating that oxidative stress might be associated with the harmful effect of autophagy suppression.

自噬是一种溶酶体降解机制，用于消除和回收受损的细胞内细胞器和蛋白质。最近的研究表明，自噬可以通过去除氧化的蛋白质和受损的线粒体来帮助减少氧化应激。自噬缺陷与许多细胞内生物过程的破坏有关。使用生物信息学工具和成纤维细胞免疫染色技术，我试图研究氧化应激是否参与介导自噬抑制对某些细胞生物学过程和信号通路的影响。许多药物成分具有不同的作用模式来抑制自噬。在这项研究中，我对中和溶酶体 pH 值（NH ₄Cl 和巴弗洛霉素 A1)。GEO 数据（GSE60570 登录号）的生物信息学分析表明，p38 信号传导诱导和 DNA 损伤反应是巴弗洛霉素 A1 处理的 RPE-1 细胞中被破坏的主要信号通路之一。同样，成纤维细胞免疫染色显示氯化铵（NH ₄Cl) 显着增加了 P38 信号、DNA 损伤标记 (H2A.X) 和氧化应激标记 (二酪氨酸)。因此，我研究了氧化应激的作用以及抗氧化剂治疗是否可以逆转自噬抑制对 p38 信号传导和 DNA 损伤反应的影响。重要的是，抗氧化处理明显恢复了自噬抑制的成纤维细胞中的 P38 信号和 H2A.X 水平。表明氧化应激可能与自噬抑制的有害影响有关。