器官移植2026，Vol.17Issue(3)：413-421,9.DOI:10.12464/j.issn.1674-7445.2026020
SPP1+巨噬细胞在慢性移植肾纤维化形成中的作用及机制研究
Study on the role and mechanism of SPP1+macrophages in the formation of chronic renal allograft fibrosis
摘要
Abstract
Objective To investigate the role and potential mechanism of secreted phosphoprotein 1(SPP1)+macrophages in the formation of chronic renal allograft fibrosis.Methods The expression features of SPP1+macrophages in renal allografts of chronic allograft dysfunction(CAD)patients were analyzed based on single-cell transcriptome data of renal tissues from patients with CAD.Transcription factor VIPER analysis and DoRothEA transcription factor activity analysis were performed on the single-cell transcriptome data.Renal tissue samples were collected from kidney transplant recipients,including the CAD group(n=5)and the non-renal allograft fibrosis group(CTL group,n=5).A mouse model of chronic allograft rejection was established and divided into the allogeneic kidney transplantation group(CAD group,n=3)and the syngeneic kidney transplantation group(SYN group,n=3).Hematoxylin-eosin staining was used to detect renal tissue injury in mice,and Masson staining was used to detect renal tissue fibrosis.Immunofluorescence staining was performed to detect SPP1 expression in renal tissues of transplant recipients and mouse renal allografts.Bone marrow-derived macrophages(BMDMs)were extracted from mice and subjected to hypoxia stimulation.The expression of hypoxia-inducible factor(HIF)-1α and SPP1 was detected by Western blot,and SPP1 expression was detected by flow cytometry.BMDMs were transfected with HIF-1α overexpression plasmid and HIF-1α small interfering RNA(siRNA)followed by hypoxia intervention,and the expression of HIF-1α and SPP1 was detected by Western blot.Mouse aortic endothelial cells(MAECs)were co-cultured with the supernatant of BMDMs,and the expression of endothelial-mesenchymal transition(EndMT)-related markers was detected by Western blot and immunofluorescence.Results Single-cell transcriptome analysis showed that the proportion of SPP1+macrophages in renal allograft tissues was significantly higher in the CAD group than in the CTL group(P＜0.05).The renal injury score and the percentage of interstitial fibrotic area in the CAD group were significantly higher than those in the SYN group(both P＜0.05).Immunofluorescence staining showed that the proportion of SPP1+macrophages was increased in the CAD group compared with the CTL group,and also increased in the CAD group compared with the SYN group(both P＜0.05).VIPER analysis and DoRothEA transcription factor activity analysis revealed activation of the hypoxia pathway and upregulated expression of transcription factors such as HIF-1α in SPP1+macrophages.SPP1 expression was elevated in BMDMs under hypoxic conditions.Knockdown of HIF-1α inhibited hypoxia-induced SPP1 protein expression,whereas overexpression of HIF-1αupregulated SPP1 protein levels.After co-culture of hypoxia-induced BMDMs with MAECs,the expression levels of EndMT-related markers were increased.Conclusions SPP1+macrophages differentiated under hypoxia are significantly infiltrated in the formation of chronic renal allograft fibrosis,and may promote renal allograft fibrosis by inducing EndMT in renal vascular endothelial cells.关键词
肾移植/纤维化/分泌型磷蛋白1/巨噬细胞/缺氧诱导因子-1α/内皮-间充质转化/内皮细胞/慢性移植肾失功Key words
Kidney Transplantation/Fibrosis/Secreted phosphoprotein 1/Macrophage/Hypoxia-inducible factor-1 α/Endothelial-mesenchymal transition/Endothelial cell/Chronic allograft dysfunction分类
医药卫生引用本文复制引用
杨泽昕,桂泽平,张俊麒,张罡,陈浩,孙黎,费爽,顾民,王子杰..SPP1+巨噬细胞在慢性移植肾纤维化形成中的作用及机制研究[J].器官移植,2026,17(3):413-421,9.基金项目
国家自然科学基金(82470790、82170769) （82470790、82170769）
