Zhang Nan-Nan, Zhu Yi, Huang Qiu-Yan, Hu Fei, Li Jun, Yang Xia
Department of Basic Medical Science, Guizhou University of Traditional Chinese Medicine, Guiyang, 550025, China.
Department of Pharmacy, Guizhou University of Traditional Chinese Medicine, Guiyang, 550025, China.
Curr Med Sci. 2025 Jul 21. doi: 10.1007/s11596-025-00091-7.
Immune infiltration and ferroptosis play pivotal roles in the progression of diabetic kidney disease (DKD). However, investigations of immune cell-related ferroptosis genes (ICRFGs) in the context of DKD are insufficient. This study aimed to identify ICRFGs relevant to DKD and screen related inhibitors.
In this study, two DKD datasets from the GEO database were utilized. We adopted the ESTIMATE algorithm to generate microenvironment scores. The CIBERSORT and WGCNA methods were employed to identify immune-related differentially expressed genes (DEGs). The common ICRFGs were derived through a Venn diagram. We employed random forest, LASSO, K-M survival, receiver operating characteristic (ROC) curve, clinical relevance, and Spearman correlation analyses to select hub ICRFGs further. Immunohistochemical experiments were also performed to validate the expression. Additionally, we utilized the Selleck database to obtain ferroptosis-related compounds and used USCF Chimera 1.14 to minimize energy, combined with molecular dynamics (MD) simulations to explore possible ferroptosis inhibitors.
Immunohistochemical analysis revealed that arachidonate 5-lipoxygenase (ALOX5) was significantly highly expressed in the db/db group. Clinical correlation and K-M survival analyses confirmed ALOX5 as the most crucial ICRFG in DKD. Furthermore, ALOX5 was significantly enriched in the terms ECM-receptor interaction, regulation of chemokine production, and regulation of the inflammatory response. A positive correlation was observed between ALOX5 and M1 macrophages, γδ T cells, and monocytes. Moreover, virtual screening and MD revealed NSC348884, salvianolic acid B, and deltarasin as potential ferroptosis inhibitors in combination with ALOX5.
We identified ALOX5 as a reliable and prospective diagnostic marker associated with immunity and ferroptosis in DKD patients.
免疫浸润和铁死亡在糖尿病肾病（DKD）进展中起关键作用。然而，在DKD背景下对免疫细胞相关铁死亡基因（ICRFGs）的研究尚不充分。本研究旨在鉴定与DKD相关的ICRFGs并筛选相关抑制剂。
本研究使用了来自GEO数据库的两个DKD数据集。我们采用ESTIMATE算法生成微环境评分。运用CIBERSORT和WGCNA方法鉴定免疫相关差异表达基因（DEGs）。通过维恩图得出共同的ICRFGs。我们采用随机森林、LASSO、K-M生存、受试者工作特征（ROC）曲线、临床相关性和Spearman相关性分析进一步选择核心ICRFGs。还进行了免疫组化实验以验证表达。此外，我们利用Selleck数据库获取铁死亡相关化合物，并使用USCF Chimera 1.14使能量最小化，结合分子动力学（MD）模拟来探索可能的铁死亡抑制剂。
免疫组化分析显示，花生四烯酸5-脂氧合酶（ALOX5）在db/db组中显著高表达。临床相关性和K-M生存分析证实ALOX5是DKD中最关键的ICRFG。此外，ALOX5在细胞外基质-受体相互作用、趋化因子产生的调节和炎症反应的调节等方面显著富集。观察到ALOX5与M1巨噬细胞、γδT细胞和单核细胞之间呈正相关。此外，虚拟筛选和MD显示NSC348884、丹酚酸B和deltarasin与ALOX5联合时为潜在的铁死亡抑制剂。
我们鉴定出ALOX5是与DKD患者免疫和铁死亡相关的可靠且有前景的诊断标志物。
