Jiang Guangwei, Li Jikuan, Dong Ruoyu, Chen Yuyan, Zhang Xiaoyu, Shi Xiaoming
Department of Vascular Surgery, Hebei General Hospital, Shijiazhuang, Hebei Province, 050000, People's Republic of China.
Second Department of Rehabilitation Medicine, The Second Hospital of Hebei Medical University, Shijiazhuang, Hebei Province, 050051, People's Republic of China.
Int J Gen Med. 2025 Jul 17;18:3969-3981. doi: 10.2147/IJGM.S522363. eCollection 2025.
During acute kidney injury, hypoxic injury following reactive oxygen species (ROS) production due to reoxidation leads to severe inflammation and ferroptosis. Thus, the aim of the current research was to determine the ferroptosis-related biomarkers in IRI injury.
GSE43974 dataset was analyzed to identify differentially expressed genes (DEGs) using bioinformatics analysis. The intersection of DEGs and ferroptosis-related genes was identified as differentially expressed ferroptosis-related genes (DEFRGs). Finally, a renal ischemia-reperfusion model was made using mice, and the model was identified by HE staining and markers of IRI, including BUN and Scr. Changes in the expression of hub gene in the model and sham groups were detected by RT-pcr.
A total of 3950 DEGs were identified between the IRI and control samples. Thereafter, 74 DEFRGs are obtained by taking the intersection of DEGs and ferroptosis-related genes. The GO analysis indicated that DEFRGs were mainly enriched in response to oxidative stress-related pathway. By MCODE, ATF3, ATF4, ATG3, ATG5, BECN1, DDIT3, HSPA5, NFE2L2, WIPI1, XBP1 were identified as hub genes. ATF3, DDIT3, ATF4, and ATG3 with AUC more than 0.7 were identified as biomarkers. It was confirmed by RT-pcr that the expression of hub genes ATF3, DDIT3, ATF4 was significantly elevated, and the expression of ATG3 was significantly reduced in the IRI model group. It was consistent with the expected results of data analysis in GEO.
In conclusion, our study identified 4 ferroptosis-related hub genes in the IRI and demonstrated that they are potential diagnostic biomarkers for IRI.
在急性肾损伤期间，由于再氧化产生的活性氧（ROS）导致的缺氧损伤会引发严重炎症和铁死亡。因此，本研究的目的是确定缺血再灌注损伤（IRI）中与铁死亡相关的生物标志物。
使用生物信息学分析对GSE43974数据集进行分析，以鉴定差异表达基因（DEG）。将DEG与铁死亡相关基因的交集鉴定为差异表达的铁死亡相关基因（DEFRG）。最后，使用小鼠建立肾缺血再灌注模型，并通过苏木精-伊红（HE）染色以及IRI标志物（包括血尿素氮（BUN）和血清肌酐（Scr））对模型进行鉴定。通过逆转录-聚合酶链反应（RT-pcr）检测模型组和假手术组中枢纽基因的表达变化。
在IRI样本与对照样本之间共鉴定出3950个DEG。此后，通过取DEG与铁死亡相关基因的交集获得74个DEFRG。基因本体（GO）分析表明，DEFRG主要富集于对氧化应激相关途径的反应。通过分子复杂检测（MCODE），鉴定出激活转录因子3（ATF3）、激活转录因子4（ATF4）、自噬相关蛋白3（ATG3）、自噬相关蛋白5（ATG5）、酵母自噬相关蛋白1（BECN1）、DNA损伤诱导转录因子3（DDIT3）、热休克蛋白家族A成员5（HSPA5）、核因子E2相关因子2（NFE2L2）、自噬相关蛋白18（WIPI1）、X盒结合蛋白1（XBP1）为枢纽基因。将曲线下面积（AUC）大于0.7的ATF3、DDIT3、ATF4和ATG3鉴定为生物标志物。通过RT-pcr证实，IRI模型组中枢纽基因ATF3、DDIT3、ATF4的表达显著升高，而ATG3的表达显著降低。这与基因表达综合数据库（GEO）中的数据分析预期结果一致。
总之，我们的研究在IRI中鉴定出4个与铁死亡相关的枢纽基因，并证明它们是IRI潜在的诊断生物标志物。
