Abstract：
Objective To elucidate the role of chaperone-mediated autophagy (CMA) in alleviating emotional dysfunction in mice with sepsis-associated encephalopathy (SAE). Methods The SAE mouse model was established by cecal ligation and perforation (CLP). The severity of sepsis was assessed using the sepsis severity score (MSS). Emotional function in SAE mice was assessed by the open-field test and elevated plus-maze. The expression levels of cognitive heat shock cognate protein 70 (HSC70), lysosomal-associated membrane protein 2A (LAMP2A) and high mobility group box 1 protein B1 (HMGB1) were detected using Western blotting. Co-localization of LAMP2A in the hippocampal neurons was observed by immunofluorescence. The release of inflammatory factors interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α) was measured using ELISA. Following 12 hours post-CLP, mice were orally administered resveratrol at a dose of 30 mg/kg once daily until day 14. Results The mortality rate of CLP mice was 45.83% 24 days post CLP, and all surviving mice exhibited emotional disturbances. 24 hours after CLP, a significant decrease in HSC70 and LAMP2A expression in hippocampal neurons was observed, indicating impaired CMA activity. Meanwhile, HMGB1 and inflammatory cytokines (IL-6 and TNF-α) levels increased. After resveratrol treatment, an increase of HSC70 and LAMP2A expression, and a decrease of HMGB1 expression and inflammatory cytokine release were observed, suggesting enhanced CMA activity and reduced neuroinflammation. Behavioral tests showed that emotional dysfunction was improved in SAE mice after resveratrol treatment. Conclusion CMA activity of hippocampal neurons in SAE mice is significantly reduced, leading to emotional dysfunction. Resveratrol can alleviate neuroinflammation and emotional dysfunction in SAE mice by promoting CMA and inhibiting the expression of HMGB1 and the release of inflammatory factors.
摘要：
目的探讨伴侣介导的自噬(CMA)在减轻脓毒症相关性脑病(SAE)小鼠情绪障碍中的作用.方法采用盲肠结扎穿孔法(CLP)建立SAE小鼠模型。使用脓毒症严重程度评分(MSS)评估脓毒症的严重程度。SAE小鼠的情绪功能通过开放视野测试和高架迷宫评估。认知热休克同源蛋白70（HSC70）的表达水平，使用蛋白质印迹法检测溶酶体相关膜蛋白2A（LAMP2A）和高迁移率族蛋白B1（HMGB1）。通过免疫荧光观察到LAMP2A在海马神经元中的共定位。ELISA法检测炎症因子白细胞介素6(IL-6)和肿瘤坏死因子α(TNF-α)的释放。CLP后12小时，小鼠每天一次以30mg/kg的剂量口服施用白藜芦醇直至第14天。结果CLP后24天小鼠死亡率为45.83%,所有幸存的小鼠都表现出情绪障碍。CLP后24小时，海马神经元中HSC70和LAMP2A表达显著下降，表明CMA活动受损。同时,HMGB1和炎性细胞因子（IL-6和TNF-α）水平升高。白藜芦醇治疗后，HSC70和LAMP2A表达增加，并观察到HMGB1表达和炎症细胞因子释放的减少，提示CMA活性增强，神经炎症减少。行为学实验表明，白藜芦醇治疗后，SAE小鼠的情绪功能障碍得到改善。结论SAE小鼠海马神经元CMA活性明显降低,导致情绪障碍。白藜芦醇可通过促进CMA、抑制HMGB1的表达和炎症因子的释放来减轻SAE小鼠的神经炎症和情绪障碍。
