Zhigao Hu , Shanshan Jiang , Zhen Wan , Laihui Luo , Minglong Wang , Hua Qiu , Yanqiang Wang , Yu Liu , Renfeng Shan
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The mRNA and protein expression levels were analyzed by RT-qPCR and western blotting, respectively. Secretion levels of pro-inflammatory cytokines were determined by ELISA. Immunofluorescence staining was employed to detect C5b-9 and LC3 levels, as well as FOXO3 cellular localization. The interaction between FOXO3 and the LAMP2 promoter was analyzed using the ChIP assay.</div></div><div><h3>Results</h3><div>MAC inhibition reduced liver injury, lipid accumulation, and inflammation in the liver tissues of ALD mice while promoting hepatocyte autophagy. CD59 overexpression not only inhibited EtOH-induced lipid accumulation and inflammation in AML12 cells but also promoted autophagy by activating the SIRT1-FOXO3 axis. Mechanistically, SIRT1 promoted FOXO3-mediated LAMP2 transcriptional activation by enhancing the deacetylation and nuclear translocation of FOXO3. As expected, SIRT1 silencing weakened the effects of CD59 overexpression on lipid accumulation, inflammatory response, and autophagy in EtOH-treated AML12 cells.</div></div><div><h3>Conclusion</h3><div>MAC inhibition enhances LAMP2-mediated hepatocyte autophagy in ALD by promoting SIRT1-mediated FOXO3 deacetylation and nuclear translocation, thereby alleviating ALD progression.</div></div>","PeriodicalId":12227,"journal":{"name":"Experimental cell research","volume":"451 2","pages":"Article 114713"},"PeriodicalIF":3.5000,"publicationDate":"2025-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Membrane attack complex impairs hepatocyte autophagy in alcohol-related liver disease by regulating the SIRT1-FOXO3 signaling\",\"authors\":\"Zhigao Hu , Shanshan Jiang , Zhen Wan , Laihui Luo , Minglong Wang , Hua Qiu , Yanqiang Wang , Yu Liu , Renfeng Shan\",\"doi\":\"10.1016/j.yexcr.2025.114713\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Background</h3><div>Impaired hepatocyte autophagy is a key feature of alcohol-related liver disease (ALD). 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引用次数: 0
摘要
背景:肝细胞自噬受损是酒精相关性肝病(ALD)的一个关键特征。激活膜攻击复合物(MAC)调节自噬。本研究探讨了MAC在ALD进展过程中肝细胞自噬中的作用和调节机制。方法采用含5%乙醇(EtOH) (w/v)的Lieber-DeCarli液体日粮建立C57BL/6小鼠模型。建立细胞模型,将AML12细胞暴露于100 mM EtOH下72 h,采用苏木精-伊红染色检测肝脏病理变化,油红O染色评估肝脏脂肪变性。RT-qPCR和western blotting分别分析mRNA和蛋白的表达水平。ELISA法检测促炎细胞因子分泌水平。免疫荧光染色检测C5b-9、LC3水平及FOXO3细胞定位。利用ChIP分析FOXO3与LAMP2启动子之间的相互作用。结果mac抑制可减轻ALD小鼠的肝损伤、脂质积累和肝组织炎症,促进肝细胞自噬。CD59过表达不仅抑制etoh诱导的AML12细胞脂质积累和炎症,而且通过激活SIRT1-FOXO3轴促进自噬。在机制上,SIRT1通过增强FOXO3的去乙酰化和核易位来促进FOXO3介导的LAMP2转录激活。正如预期的那样,SIRT1沉默减弱了CD59过表达对etoh处理的AML12细胞中脂质积累、炎症反应和自噬的影响。结论mac抑制通过促进sirt1介导的FOXO3去乙酰化和核易位,增强了lamp2介导的ALD肝细胞自噬,从而缓解ALD的进展。
Membrane attack complex impairs hepatocyte autophagy in alcohol-related liver disease by regulating the SIRT1-FOXO3 signaling
Background
Impaired hepatocyte autophagy is a key feature of alcohol-related liver disease (ALD). Activation of the membrane attack complex (MAC) regulates autophagy. This study examined the role and regulatory mechanisms of MAC in hepatocyte autophagy during ALD progression.
Methods
For the animal model, C57BL/6 mice were fed a Lieber-DeCarli liquid diet containing 5 % ethanol (EtOH) (w/v). For the cell model, AML12 cells were exposed to 100 mM EtOH for 72 h. Pathological changes in the liver were examined using hematoxylin–eosin staining, and hepatic steatosis was evaluated using Oil Red O staining. The mRNA and protein expression levels were analyzed by RT-qPCR and western blotting, respectively. Secretion levels of pro-inflammatory cytokines were determined by ELISA. Immunofluorescence staining was employed to detect C5b-9 and LC3 levels, as well as FOXO3 cellular localization. The interaction between FOXO3 and the LAMP2 promoter was analyzed using the ChIP assay.
Results
MAC inhibition reduced liver injury, lipid accumulation, and inflammation in the liver tissues of ALD mice while promoting hepatocyte autophagy. CD59 overexpression not only inhibited EtOH-induced lipid accumulation and inflammation in AML12 cells but also promoted autophagy by activating the SIRT1-FOXO3 axis. Mechanistically, SIRT1 promoted FOXO3-mediated LAMP2 transcriptional activation by enhancing the deacetylation and nuclear translocation of FOXO3. As expected, SIRT1 silencing weakened the effects of CD59 overexpression on lipid accumulation, inflammatory response, and autophagy in EtOH-treated AML12 cells.
Conclusion
MAC inhibition enhances LAMP2-mediated hepatocyte autophagy in ALD by promoting SIRT1-mediated FOXO3 deacetylation and nuclear translocation, thereby alleviating ALD progression.
期刊介绍:
Our scope includes but is not limited to areas such as: Chromosome biology; Chromatin and epigenetics; DNA repair; Gene regulation; Nuclear import-export; RNA processing; Non-coding RNAs; Organelle biology; The cytoskeleton; Intracellular trafficking; Cell-cell and cell-matrix interactions; Cell motility and migration; Cell proliferation; Cellular differentiation; Signal transduction; Programmed cell death.