高糖环境下Z-DNA结合蛋白1调控巨噬细胞铁死亡的转录组学机制研究OA

Objective To investigate the role of Z-DNA binding protein 1(ZBP1)in regulating ferroptosis in macrophages under high-glucose conditions and its potential molecular mechanisms.Methods The murine macrophage cell line J774A.1 was used as the research object.A cell injury model was established using high glucose(40 mmol/L glucose),and a ZBP1 stable knockdown cell line was constructed via short hairpin RNA(shRNA).qRT-PCR was performed to detect ZBP1 expres-sion levels to verify the efficacy of model construction.Transcriptome sequencing was conducted on high-glucose group(HG)and ZBP1-knockdown high-glucose group(HG+shZBP1),and differentially expressed genes were screened.Gene ontology(GO),Kyoto encyclopedia of genes and genomes(KEGG),gene set enrichment analysis(GSEA),and protein-protein inter-action(PPI)network analyses were subsequently performed.Ferroptosis-related differentially expressed genes were selected for qRT-PCR validation.Meanwhile,intracellular reactive oxygen species(ROS)levels were detected by flow cytometry,and total iron content and reduced glutathione(GSH)content were measured.Results ZBP1 expression was significantly upregulated by high-glucose stimulation,and was decreased by approximately 89％after ZBP1 knockdown compared with the control group,indicating that the knockdown model was successfully constructed.As revealed by transcriptome sequencing,a total of 710 dif-ferentially expressed genes(384 upregulated and 326 downregulated)were screened in the HG+shZBP1 group compared with the HG group.GO,KEGG,and GSEA analyses indicated that the differential genes were mainly enriched in redox regulation,inflammatory response,iron ion binding,lipid metabolism,and MAPK signaling pathways.A close interaction among ferropto-sis-related genes was revealed by PPI network analysis.qRT-PCR validation results showed that Gpx4 expression was signifi-cantly upregulated,whereas Alox15 and Tfrc expressions were significantly downregulated in the HG+shZBP1 group compared with the HG group.Meanwhile,ROS levels and total iron content were significantly decreased,and GSH content was signifi-cantly increased in the HG+shZBP1 group(n=3,P<0.05).Conclusion High-glucose conditions can induce oxidative stress,iron metabolism dysregulation,and abnormal expression of ferroptosis-related genes in macrophages,accompanied by upregulated ZBP1 expression.ZBP1 knockdown ameliorates high-glucose-induced oxidative stress,mitigates iron metabolism imbalance,and modulates the expression of ferroptosis-related genes,suggesting that ZBP1 may be involved in the regulation of macrophage ferroptosis under high-glucose conditions.However,these findings require further validation through in vivo studies and clinical samples.