早期生长应答因子1对THP-1源性泡沫细胞铜死亡相关基因表达的影响及其机制OA

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.