Zhang Zhuohua, Liu Yan, Huang Da, Huang Zhaohe
Department of Cardiology, First Affiliated Hospital of Jinan University, Guangzhou, 510630, People's Republic of China.
Department of Cardiology, Affiliated Hospital of Youjiang Medical University for Nationalities, Baise, 533000, People's Republic of China.
J Inflamm Res. 2024 Sep 9;17:6203-6227. doi: 10.2147/JIR.S476456. eCollection 2024.
Myocardial ischemia-reperfusion injury (MIRI) is characterized by inflammation and ferroptosis, but the precise mechanisms remain unknown. This study used single-cell transcriptomics technology to investigate the changes in various cell subtypes during MIRI and the regulatory network of ferroptosis-related genes and immune infiltration.
Datasets GSE146285, GSE83472, GSE61592, and GSE160516 were obtained from Gene Expression Omnibus. Each cell subtype in the tissue samples was documented. The Seurat package was used for data preprocessing, standardization, and clustering. Cellphonedb was used to investigate the ligand-receptor interactions between cells. The hdWGCNA analysis was used to create a gene co-expression network. GSVA and GSEA were combined to perform functional enrichment and pathway analysis on the gene set. Furthermore, characteristic genes of the disease were identified using Lasso regression and SVM algorithms. Immune cell infiltration analysis was also performed. MIRI rat models were created, and samples were taken for RT-qPCR and Western blot validation.
The proportion of MIRI samples in the C2, C6, and C11 subtypes was significantly higher than that of control samples. Three genes associated with ferroptosis (CD44, Cfl1, and Zfp36) were identified as MIRI core genes. The expression of these core genes was significantly correlated with mast cells and monocyte immune infiltrating cells. The experimental validation confirmed the upregulation of Cd44 and Zfp36 expression levels in MIRI, consistent with current study trends.
This study used single-cell transcriptomics technology to investigate the molecular mechanisms underpinning MIRI. Numerous important cell subtypes, gene regulatory networks, and disease-associated immune infiltration were also discovered. These findings provide new information and potential therapeutic targets for MIRI diagnosis and treatment.
心肌缺血再灌注损伤（MIRI）以炎症和铁死亡为特征，但其确切机制仍不清楚。本研究采用单细胞转录组学技术，研究MIRI过程中各种细胞亚型的变化以及铁死亡相关基因的调控网络和免疫浸润情况。
从基因表达综合数据库（Gene Expression Omnibus）获取数据集GSE146285、GSE83472、GSE61592和GSE160516。记录组织样本中的每种细胞亚型。使用Seurat软件包进行数据预处理、标准化和聚类。使用Cellphonedb研究细胞间的配体-受体相互作用。使用hdWGCNA分析创建基因共表达网络。结合基因集变异分析（GSVA）和基因集富集分析（GSEA）对基因集进行功能富集和通路分析。此外，使用套索回归和支持向量机算法鉴定疾病特征基因。还进行了免疫细胞浸润分析。建立MIRI大鼠模型，并采集样本进行逆转录定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹（Western blot）验证。
C2、C6和C11亚型中MIRI样本的比例显著高于对照样本。鉴定出三个与铁死亡相关的基因（CD44、Cfl1和Zfp36）为MIRI核心基因。这些核心基因的表达与肥大细胞和单核细胞免疫浸润细胞显著相关。实验验证证实了MIRI中Cd44和Zfp36表达水平上调，与当前研究趋势一致。
本研究采用单细胞转录组学技术研究MIRI的分子机制。还发现了许多重要的细胞亚型、基因调控网络和疾病相关的免疫浸润。这些发现为MIRI的诊断和治疗提供了新的信息和潜在的治疗靶点。
