江苏大学学报（医学版）2025，Vol.35Issue(4)：277-284,8.DOI:10.13312/j.issn.1671-7783.y240127
脂毒性肝细胞外泌体通过抑制铁死亡促进肝星状细胞活化
Lipotoxic hepatocyte exosomes promote hepatic stellate cell activation by inhibiting ferroptosis
摘要
Abstract
Objective:To investigate the impact of lipotoxic hepatocyte-derived exosome(LTH-ex)on ferroptosis and activation of hepatic stellate cell(HSC).Methods:Lipotoxic damage was induced in the human liver cell line LO2 using oleic acid and palmitic acid.The supernatants of the lipotoxic damaged liver cells were collected and LTH-ex was extracted through ultracentrifugation.The characterization of LTH-ex was identified using transmission electron microscopy and Nanosight nanoparticle analysis system,including assessing morphology,particle size and concentration,and LTH-ex surface markers through Western blotting.The human HSC cell line LX-2 was co-cultured with LTH-ex or the ferroptosis activator Erastin for 48 h,and then all cells were divided into three groups:the PBS control group,the LTH-ex treatment group,and the co-treatment group consisting of LTH-ex and the ferroptosis inducer Erastin.Cell viability and mitochondrial membrane potential of LX-2 were evaluated using FDA and JC-1 staining.The iron ion concentration kit was used to detect the iron level in LX-2,the expression of mRNA and protein of death suppressor genes GPX4,xCT,and HSC activation gene α-SMA,were measured through qRT-PCR and Western blotting.Additionally,a mouse model of non-alcoholic fatty liver disease was established by feeding a high-fat diet,followed by injection of LTH-ex through the tail vein.Liver tissue structure was observed using HE staining,while GPX4 and α-SMA protein expression was assessed through immunohistochemistry staining.Sirius red staining was employed to detect collagen deposition in the liver tissue.Results:The LTH-ex particles were approximately 110 nm in size,displaying the typical lipid bilayer structure of exosomes and containing exosome marker proteins TSG101,Alix,and CD63.In comparison to the PBS control group,the LTH-ex treatment group demonstrated enhanced viability of LX-2 cells,an increased mitochondrial membrane potential,decreased iron levels,and upregulation of the ferroptosis-inhibiting genes GPX4 and xCT,as well as HSC activation gene α-SMA.Conversely,when compared to the LTH-ex treatment group,the co-treatment group of LTH-ex and the ferroptosis inducer Erastin exhibited reduced LX-2 cell viability and mitochondrial membrane potential,alongside downregulation of GPX4,xCT,and α-SMA.HE staining,immunohistochemistry,and Sirius red staining demonstrated that LTH-ex could exacerbate vacuolar degeneration of hepatocytes in the liver tissue of mice with non-alcoholic fatty liver disease.Additionally,LTH-ex promoted the expression of GPX4 and α-SMA proteins and increased collagen deposition in the liver tissue.expression.Conclusion:LTH-ex increase the cell viability of hepatic stellate cells and the expression of ferroptosis inhibitory genes GPX4 and xCT,and it may be implicated that LTH-ex may promote hepatic stellate cell activation by inhibiting ferroptosis,and exacerbate collagen deposition in the liver tissue of non-alcoholic fatty liver disease mice.关键词
脂毒性肝细胞/外泌体/铁死亡/肝星状细胞活化/非酒精性脂肪性肝病Key words
lipotoxic hepatocytes/exosome/ferroptosis/hepatic stellate cell activation/nonalcoholic fatty liver disease分类
医药卫生引用本文复制引用
史雨杰,王岩金,严永敏..脂毒性肝细胞外泌体通过抑制铁死亡促进肝星状细胞活化[J].江苏大学学报（医学版）,2025,35(4):277-284,8.基金项目
国家自然科学基金面上项目(82272421) （82272421）
常州市科技支撑计划项目(CE20235004) （CE20235004）
