Wang Qin, Liu Yuxing, Zhang Yan, Zhang Siyuan, Zhao Meifang, Peng Zhangzhe, Xu Hui, Huang Hao
Department of Nephrology, Xiangya Hospital, Central South University, Changsha, China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, China.
Department of Nephrology, Xiangya Hospital, Central South University, Changsha, China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, China; Department of Cell Biology, School of Life Sciences, Central South University, Changsha, China.
Int Immunopharmacol. 2024 Mar 30;130:111754. doi: 10.1016/j.intimp.2024.111754. Epub 2024 Mar 1.
Acute kidney injury (AKI) is a complex disease, with macrophages playing a vital role in its progression. However, the mechanism of macrophage function remains unclear and strategies targeting macrophages in AKI are controversial. To address this issue, we used single-cell RNA-seq analysis to identify macrophage sub-types involved in ischemia-reperfusion-induced AKI, and then screened for associated hub genes using intersecting bulk RNA-seq data. The single-cell and bulk RNA-seq datasets were obtained from the Gene Expression Omnibus (GEO) database. Screening of differentially-expressed genes (DEGs) and pseudo-bulk DEG analyses were used to identify common hub genes. Pseudotime and trajectory analyses were performed to investigate the progression of cell differentiation. CellChat analysis was performed to reveal the crosstalk between cell clusters. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were used to identify enriched pathways in the cell clusters. Immunofluorescence and RT-PCR were preformed to validate the expression of the identified hub genes. Four hub genes, Vim, S100a6, Ier3, and Ccr1, were identified in the infiltrated macrophages between normal samples and those 3 days after ischemia-reperfusion renal injury (IRI); all were associated with the progression of IRI-induced AKI. Increased expression of Vim, S100a6, Ier3, and Ccr1 in infiltrated macrophages may be associated with inflammatory responses and may mediate crosstalk between macrophages and renal tubular epithelial cells under IRI conditions. Our results reveal that Ier3 may be critical in AKI, and that Vim, S100a6, Ier3, and Ccr1 may act as novel biomarkers and potential therapeutic targets for IRI-induced AKI.
急性肾损伤（AKI）是一种复杂的疾病，巨噬细胞在其进展过程中起着至关重要的作用。然而，巨噬细胞功能的机制仍不清楚，针对AKI中巨噬细胞的策略也存在争议。为了解决这个问题，我们使用单细胞RNA测序分析来鉴定参与缺血再灌注诱导的AKI的巨噬细胞亚型，然后使用交叉的批量RNA测序数据筛选相关的核心基因。单细胞和批量RNA测序数据集来自基因表达综合数据库（GEO）。通过筛选差异表达基因（DEG）和伪批量DEG分析来鉴定常见的核心基因。进行伪时间和轨迹分析以研究细胞分化的进程。进行CellChat分析以揭示细胞簇之间的串扰。使用基因本体论和京都基因与基因组百科全书分析来鉴定细胞簇中富集的途径。进行免疫荧光和RT-PCR以验证所鉴定的核心基因的表达。在正常样本与缺血再灌注肾损伤（IRI）后3天的样本之间浸润的巨噬细胞中鉴定出四个核心基因，即波形蛋白（Vim）、S100钙结合蛋白A6（S100a6）、早期反应基因3（Ier3）和C趋化因子受体1（Ccr1）；所有这些基因都与IRI诱导的AKI的进展相关。浸润的巨噬细胞中Vim、S100a6、Ier3和Ccr1表达的增加可能与炎症反应相关，并且可能在IRI条件下介导巨噬细胞与肾小管上皮细胞之间的串扰。我们的结果表明，Ier3可能在AKI中起关键作用，并且Vim、S100a6、Ier3和Ccr1可能作为IRI诱导的AKI的新型生物标志物和潜在治疗靶点。
