摘要
目的探讨甘草查尔酮A(LA)对H3N2亚型甲型流感病毒(IAV)感染巨噬细胞损伤的保护作用,并阐明其调控铁死亡的作用机制。方法建立IAV H3N2感染RAW264.7细胞模型。实验分为对照组、IAV模型组、IAV+LA组、IAV+LA+LY294002组、IAV+LA+OE-p53组。采用CCK-8法检测细胞活力;qPCR和Western Blot检测铁死亡相关分子(TFR1、FTH1、GPX4、p53)的表达;流式细胞术检测活性氧(ROS)水平;ELISA法检测炎症因子(IL-6、TNF-α、IL-1β)含量;试剂盒检测MDA、Fe^(2+)和GSH水平。结果IAV感染诱导铁死亡:病毒显著抑制细胞活力(降至58.3%),上调ROS、炎症因子、MDA、Fe^(2+)、TFR1及p53水平,同时下调GSH、GPX4和FTH1表达。LA的保护作用:LA干预逆转上述变化,提高细胞活力,抑制氧化应激与炎症,并调节铁代谢稳态(如降低TFR1、提升FTH1和GPX4)。机制依赖p53/PI3K通路:使用PI3K抑制剂LY294002或过表达p53均削弱LA对铁死亡指标(MDA、Fe^(2+)、GSH、TFR1/FTH1/GPX4)的调控,同时,也削弱LA对ROS和炎症因子的抑制作用,表明LA的抗铁死亡作用可能依赖于该信号轴。结论LA能够减轻H3N2流感病毒诱导的巨噬细胞损伤,其机制可能通过调控p53/PI3K信号轴,抑制TFR1介导的铁摄取、促进FTH1介导的铁储存、增强GPX4/GSH抗氧化系统,从而抑制铁死亡进程。
Objective To investigate the protective effect of Licochalcone A(LA)on H3N2 subtype influenza A virus(IAV)-induced macrophage injury and to elucidate its regulatory mechanism on ferroptosis.Methods An IAV H3N2-infected RAW264.7 macrophage model was established.Cells were divided into five groups:control,IAV model,IAV+LA,IAV+LA+LY294002,and IAV+LA+OE-p53.Cell viability was assessed using the CCK-8 assay.The expression levels of ferroptosis-related molecules(TFR1,FTH1,GPX4,p53)were measured by quantitative real-time PCR(qPCR)and Western Blot.Intracellular reactive oxygen species(ROS)levels were detected by flow cytometry.The levels of inflammatory factors(IL-6,TNF-α,IL-1β)were detected by enzyme-linked immunosorbent assay(ELISA).The levels of malondialdehyde(MDA),ferrous ion(Fe^(2+)),and glutathione(GSH)were determined using commercial assay kits.Results IAV infection induced ferroptosis,as evidenced by a significant decrease in cell viability(to 58.3%),upregulation of ROS,inflammatory cytokines,MDA,Fe^(2+),TFR1,and p53 levels,alongside downregulation of GSH,GPX4,and FTH1 expression.LA intervention reversed these alterations:it improved cell viability,suppressed oxidative stress and inflammation,and modulated iron metabolism homeostasis(e.g.,decreased TFR1,increased FTH1 and GPX4).The mechanism depended on the p53/PI3K pathway.Both the application of the PI3K inhibitor LY294002 and p53 overexpression attenuated LA's regulatory effects on ferroptosis indicators(MDA,Fe^(2+),GSH,and TFR1/FTH1/GPX4)and its inhibitory effects on ROS and inflammatory cytokines,indicating that LA's anti-ferroptosis action was likely dependent on this signaling axis.Conclusion LA can alleviate H3N2 influenza virus-induced macrophage injury.The underlying mechanism may involve the regulation of the p53/PI3K signaling axis,leading to the inhibition of TFR1-mediated iron uptake,promotion of FTH1-mediated iron storage,and enhancement of the GPX4/GSH antioxidant system,thereby suppressing the ferroptosis process.
作者
黄津
陈家卫
李声忠
黄美珍
张美泉
Huang Jin;Chen Jiawei;Li Shengzhong;Huang Meizhen;Zhang Meiquan(Department of Infectious Diseases,the Second Affiliated Hospital of Fujian University of Traditional Chinese Medicine,Fuzhou 350003;Department of Respiratory and Critical Care Medicine,Fujian Provincial Geriatric Hospital,Fuzhou 350009)
出处
《中国现代医药杂志》
2026年第3期19-25,共7页
Modern Medicine Journal of China
基金
福建省自然科学基金项目(编号:2023J01814)。
