Objective To explore the role and mechanisms of interferon regulatory factor 8(IRF8)in the pathogenesis of acute lung injury(ALI).Methods Reverse transcription-quantitative polymerase chain reaction(RT-qPCR)and western blots(WB)were used to detect the expression of IRF8 protein and mRNA in models of lung injury both in vivo and in vitro.IRF8 stable overexpression and knockdown A549 cell lines were established in vitro,and the expression levels of inflammation-related genes at the transcriptional level were detected using RT-qPCR.WB analysis was employed to assess the expression of apoptosis-related genes.Each group of 12 wild type(WT)mice and Irf8 knockout mice was selected to establish the model of ALI.Collect lung tissues from the modeled mice and weigh them.Bronchoalveolar lavage fluid(BALF)was collected and assessed for total cell count and total protein content.Hematoxylin-eosin(HE)staining and immunofluorescence(IF)analyses were performed to evaluate lung tissue inflammation infiltration.RT-qPCR was used to detect the mRNA expression of inflammation-related genes in the lung tissue of Irf8 knockout mice.The content of apoptosis in lung tissue was detected by TUNEL staining.In vivo and in vitro,the protein expression of ferroptosis-related signaling molecules was assessed by WB.Results The expression of IRF8 is upregulated both in mRNA and protein levels in ALI.IRF8 knockdown exacerbates intracellular inflammation and cell apoptosis in vitro.Conversely,IRF8 overexpression can exert a protective effect in vitro.Irf8 knockout further exacerbates lung injury in vivo.Mechanistically,IRF8 alleviates lung injury by regulating the expression of ferroptosis-related signaling molecules.Conclusion IRF8 exerts a protective role in lung injury by targeting ferroptosis,providing a potential target for the treatment of lung injury.
维普
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