In sepsis, bacterial components, particularly lipopolysaccharide (LPS), trigger organ injuries such as liver dysfunction. Although sepsis induces hepatocyte damage, the mechanisms underlying sepsis‐related hepatic failure remain unclear. In this study, we demonstrated that the LPS‐treated rat hepatocyte cell line Clone 9 not only induced reactive oxygen species (ROS) generation and apoptosis but also increased the expression of the autophagy marker proteins LC3‐II and p62, and decreased the expression of intact Lamp2A, a lysosomal membrane protein. Additionally, LPS increased lysosomal membrane permeability and galectin‐3 puncta formation, and promoted lysosomal alkalization in Clone 9 cells. Pharmacological inhibition of caspase‐8 and cathepsin D (CTSD) suppressed the activation of caspase‐3 and rescued the viability of LPS‐treated Clone 9 cells. Furthermore, LPS induced CTSD release associated with lysosomal leakage and contributed to caspase‐8 activation. Pretreatment with the antioxidant N‐acetylcysteine (NAC) not only diminished ROS generation and increased the cell survival rate, but also decreased the expression of activated caspase‐8 and caspase‐3 and increased the protein level of Lamp2A in LPS‐treated Clone 9 cells. These results demonstrate that LPS‐induced ROS causes lysosomal membrane permeabilization and lysosomal cell death, which may play a crucial role in hepatic failure in sepsis. Our results may facilitate the development of new strategies for sepsis management.

中文翻译：

---

脂多糖通过大鼠肝细胞克隆中的活性氧诱导溶酶体细胞死亡 9


在脓毒症中，细菌成分，特别是脂多糖 （LPS），会引发器官损伤，例如肝功能障碍。尽管脓毒症会诱发肝细胞损伤，但脓毒症相关肝功能衰竭的机制仍不清楚。在这项研究中，我们证明 LPS 处理的大鼠肝细胞系克隆 9 不仅诱导活性氧 （ROS） 产生和细胞凋亡，还增加了自噬标志物蛋白 LC3-II 和 p62 的表达，并降低了溶酶体膜蛋白完整 Lamp2A 的表达。此外，LPS 增加了溶酶体膜通透性和半乳糖凝集素-3 点状体形成，并促进了克隆 9 细胞中的溶酶体碱化。caspase-8 和组织蛋白酶 D （CTSD） 的药理学抑制抑制了 caspase-3 的激活并挽救了 LPS 处理的克隆 9 细胞的活力。此外，LPS 诱导与溶酶体渗漏相关的 CTSD 释放，并促进 caspase-8 激活。用抗氧化剂 N-乙酰半胱氨酸 （NAC） 预处理不仅减少了 ROS 的产生并提高了细胞存活率，而且还降低了活化的 caspase-8 和 caspase-3 的表达，并增加了 LPS 处理的克隆 9 细胞中 Lamp2A 的蛋白水平。这些结果表明，LPS 诱导的 ROS 导致溶酶体膜通透和溶酶体细胞死亡，这可能在脓毒症肝功能衰竭中起关键作用。我们的结果可能有助于开发脓毒症管理的新策略。