摘要
目的观察早期生长应答因子1(EGR1)对THP-1源性泡沫细胞铜死亡相关基因表达的影响,并探讨其机制。方法取人单核细胞系THP-1,用佛波酯(PMA)诱导分化为THP-1源性巨噬细胞后,加入氧化型低密度脂蛋白(ox-LDL)诱导分化为THP-1源性泡沫细胞,油红O染色后观察细胞形态。分别取THP-1源性巨噬细胞(对照组)、THP-1源性泡沫细胞(模型组)、转染EGR1过表达质粒LV-OE-EGR1的THP-1源性泡沫细胞(过表达组)、转染EGR1敲低质粒si-EGR1的THP-1源性泡沫细胞(敲低组)、转染EGR1敲低质粒si-EGR1并加入自噬抑制剂氯喹(CQ)的THP-1源性泡沫细胞(敲低+抑制自噬组),采用RT-qPCR法检测细胞自噬标志物[微管相关蛋白1轻链3(LC3)、p62]、铜死亡相关基因[铁氧还蛋白1(FDX1)、二氢硫辛酸转乙酰基酶(DLAT)]mRNA,采用Western blotting法检测细胞自噬标志物、铜死亡相关蛋白。结果显微镜下观察到,细胞胞质内被染为红色或橙色的脂滴,表明THP-1源性泡沫细胞模型构建成功。与对照组相比,模型组细胞LC3 mRNA和蛋白相对表达量均升高(P均<0.05),p62、FDX1、DLAT mRNA和蛋白相对表达量均降低(P均<0.05)。与模型组相比,过表达组细胞LC3 mRNA和蛋白相对表达量均降低(P均<0.05),p62、FDX1、DLAT mRNA和蛋白相对表达量均升高(P均<0.05);敲低组细胞LC3 mRNA和蛋白相对表达量均升高(P均<0.05),p62、FDX1、DLAT mRNA和蛋白相对表达量均降低(P均<0.05)。与敲低组相比,敲低+抑制自噬组细胞LC3 mRNA和蛋白相对表达量均降低(P均<0.05),p62、FDX1、DLAT mRNA和蛋白相对表达量均升高(P均<0.05)。结论EGR1可通过自噬途径调控THP-1源性泡沫细胞铜死亡相关基因FDX1、DLAT的表达。
Objective To observe the effect of early growth response factor 1(EGR1)on the expression of cupropto⁃sis-related genes in THP-1-derived foam cells and to explore its underlying mechanism.Methods The human monocyte cell line THP-1 was first differentiated into THP-1-derived macrophages using phorbol ester(PMA),followed by treatment with oxidized low-density lipoprotein(ox-LDL)to induce differentiation into THP-1-derived foam cells.Cell morphology was observed after Oil Red O staining.RT-qPCR was used to detect the mRNA expression of autophagy markers[microtu⁃bule-associated protein 1 light chain 3(LC3)and p62]and cuproptosis-related genes[ferredoxin 1(FDX1)and dihydroli⁃poamide S-acetyltransferase(DLAT)]in THP-1-derived macrophages(control group),THP-1-derived foam cells(model group),THP-1-derived foam cells transfected with the EGR1 overexpression plasmid LV-OE-EGR1(overexpression group),THP-1-derived foam cells transfected with the EGR1 knockdown plasmid si-EGR1(knockdown group),and THP-1-derived foam cells transfected with si-EGR1 and treated with the autophagy inhibitor chloroquine(CQ)(knockdown+autophagy inhibition group).Western blotting was performed to detect the protein levels of autophagy markers and cupro⁃ptosis-related proteins in the respective groups.Results Under microscopic examination,red or orange lipid droplets were observed in the cytoplasm,indicating successful construction of the THP-1-derived foam cell model.Compared with the control group,the model group showed increased relative expression levels of LC3 mRNA and protein(all P<0.05),while the relative expression levels of p62,FDX1,and DLAT mRNA and protein decreased(all P<0.05).Compared with the model group,the overexpression group exhibited decreased relative expression levels of LC3 mRNA and protein(all P<0.05)and increased relative expression levels of p62,FDX1,and DLAT mRNA and protein(all P<0.05).In contrast,the knockdown group showed increased relative expression levels of LC3 mRNA and protein(all P<0.05)and decreased relative expression levels of p62,FDX1,and DLAT mRNA and protein(all P<0.05).Compared with the knockdown group,the knockdown+autophagy inhibition group demonstrated decreased relative expression levels of LC3 mRNA and protein(all P<0.05)and increased relative expression levels of p62,FDX1,and DLAT mRNA and protein(all P<0.05).Conclusion EGR1 can regulate the expression of cuproptosis-related genes FDX1 and DLAT in THP-1-derived foam cells through the autophagy pathway.
作者
崔芸
杨娟
陈涵
王红
张黎
CUI Yun;YANG Juan;CHEN Han;WANG Hong;ZHANG Li(Ward 17,Fuwai Yunnan Hospital,Chinese Academy of Medical Sciences,Yunnan Hospital,Fuwai Hospital Chinese Academy of Medical Sciences,Affiliated Cardiovascular Hospital of Kunming Medical University,Kunming 650102,China;不详)
出处
《山东医药》
2026年第1期1-6,共6页
Shandong Medical Journal
基金
云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目(202301AY070001-204)
云南省阜外心血管病医院青年人才托举资助项目(2023RCTJ-QN001)。
