周焕杰,朱浪静

背景：脓毒症急性肾损伤（Sepsis-induced acute kidney injury，SI-AKI）是指由于严重感染导致全身炎症反应，进而引起的肾脏功能急剧下降。如果不及时治疗，可能会导致肾功能衰竭，甚至危及生命。目前，尚缺乏能够有效且便捷地提示SI-AKI疾病严重程度及临床结果的生物标志物。中性粒细胞外陷阱（Neutrophil Extracellular Traps，NETs）是由中性粒细胞释放的一种由细胞核和细胞质成分组成的网状结构，能够将病原体缠绕起来，防止它们扩散和感染。这一过程是免疫系统防御机制的一部分。目的：利用SI-AKI小鼠肾脏的单细胞数据及脓毒症AKI患者外周血转录组数据，结合机器学习和免疫浸润分析，深入分析NETs基因在SI-AKI中的作用。方法：从GEO数据库中下载并分析GSE232404数据集，该数据集包含5例SI-AKI患者和5例正常健康全血样本，同时从GeneCard数据库下载NETs基因数据。通过对获得的两组数据集筛选差异表达基因，并对其进行GO/KEGG通路分析，获得NETs相关差异表达基因。通过构建出PPI网络，并采用随机森林算法以及最小绝对收缩和选择算子机器学习技术两种算法，筛选出核心关键基因，进行诊断预测。同时采用CIBERSORT分析SI-AKI免疫浸润模式，采用Spearman法分析该核心关键基因与免疫浸润丰度的关系。此外还进行了单细胞测序分析和拟时序分析，以在单细胞水平上研究该核心关键基因在SI-AKI中的作用机制。结果：在转录组分析中，基于蛋白质-蛋白质相互作用（Protein-Protein Interaction，PPI）和机器学习分析，我们筛选出了一个与NETs相关的差异表达基因，即基质金属蛋白酶9（Matrix MetalloPeptidase 9，MMP9）。免疫学分析显示，在SI-AKI中中性粒细胞、巨噬细胞浸润显著增加。Spearman相关分析表明，MMP9与促炎免疫细胞的浸润呈正相关，但与抗炎或调节性免疫细胞的浸润呈负相关。单细胞测序显示MMP9在中性粒细胞显著高表达。结论：MMP9是SI-AKI中的关键NETs基因，并参与了SI-AKI的炎症反应，这为理解SI-AKI的发病机制提供了新的视角。

脓毒症急性肾损伤；中性粒细胞外陷阱；单细胞测序；机器学习

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