背景:糖尿病肾病(DKD)是一种常见的微血管并发症,也是糖尿病患者死亡的主要原因之一。Ferroptosis,以脂质ROS积累为特征的铁依赖性细胞死亡模式,被发现与许多疾病有关,并且在肾脏疾病中具有巨大的潜力。在DKD中鉴定潜在的铁凋亡相关基因及其生物学机制具有重要价值。
方法:我们从基因表达综合(GEO)数据库获得GSE30122数据集,并从Ferrdb数据库获得铁凋亡相关基因。差异表达分析后,和三种机器学习算法,确定了中心铁凋亡相关基因EZH2。为了研究EZH2的功能,基因集富集分析(GSEA),进行基因集变异分析(GSVA)和单细胞分析。EZH2在DKD患者中的表达得到验证,HK-2细胞模型和DKD小鼠模型。进行EZH2敲低HK-2细胞和用GSK126处理的HK-2细胞以验证EZH2是否影响DKD中的铁凋亡。CHIP测定用于检测EZH2是否通过抑制SLC7A11来调节铁凋亡。进行分子对接以探索EZH2和四种中药(SennosideA,小檗碱,伞形酮,桔梗皂苷D)与DKD治疗中的铁凋亡有关。
结果:根据GEO中的GSE30122数据集和来自Ferrb数据库的铁性凋亡相关基因,我们通过多样化的机器学习方法获得了DKD中的中枢铁死亡相关基因EZH2。在单细胞分析中显示EZH2表达的增加,DKD患者,DKD小鼠模型和高糖诱导的DKD细胞模型。进一步研究表明,EZH2敲除和抑制可以减轻体外HG诱导的铁凋亡。CHIP分析显示EZH2介导的表观遗传沉默调节SLC7A11的表达。分子对接结果表明,EZH2与人参皂甙A具有较强的结合稳定性,小檗碱,伞形酮,
结论:总的来说,我们的数据表明,组蛋白H3K27甲基转移酶EZH2可以通过抑制DKD中的SLC7A11来调节肾小管上皮细胞的铁凋亡,这可能是诊断DKD的可靠可靠指标和潜在的治疗目标。
BACKGROUND: Diabetic kidney disease (DKD) is a common microvascular complication and one of the main causes of death in diabetes. Ferroptosis, an iron-dependent mode of cell death characterized by lipid ROS accumulation, was found to be associated with a number of diseases and has great potential for kidney diseases. It has great value to identify potential ferroptosis-related genes and their biological mechanisms in DKD.
METHODS: We obtained the GSE30122 dataset from Gene Expression Omnibus (GEO) database and ferroptosis-related genes from the Ferrdb database. After differential expression analysis, and three machine learning algorithms, the hub ferroptosis-related gene EZH2 was identified. In order to investigate the function of EZH2, Gene Set Enrichment Analysis (GSEA), Gene Set Variation Analysis (GSVA) and single cell analysis were conducted. The expression of EZH2 was validated in DKD patients, HK-2 cell models and DKD mouse models. EZH2 knockdown HK-2 cells and HK-2 cells treated with GSK126 were performed to verify whether EZH2 affected ferroptosis in DKD. CHIP assay was used to detect whether EZH2 regulated ferroptosis by suppressing SLC7A11. Molecular docking was performed to explore EZH2 and four traditional Chinese medicine (Sennoside A, Berberine, Umbelliferone, Platycodin D) related to ferroptosis in DKD treatment.
RESULTS: According to the GSE30122 dataset in GEO and ferroptosis-related genes from the Ferrb database, we obtained the hub ferroptosis-related gene EZH2 in DKD via diversified machine learning methods. The increasing of EZH2 expression was shown in single cell analysis, DKD patients, DKD mouse models and high glucose induced DKD cell models. Further study showed that EZH2 knockdown and inhibition can alleviate HG-induced ferroptosis in vitro. CHIP assay showed EZH2-mediated epigenetic silencing regulated the expression of SLC7A11. Molecular docking results showed that EZH2 had strong binding stability with Sennoside A, Berberine, Umbelliferone, and Platycodin D.
CONCLUSIONS: Overall, our data shouwed that histone H3K27 methyltransferase EZH2 could regulate the renal tubular epithelial cell ferroptosis by suppressing SLC7A11 in DKD, which may serve as a credible reliable indicator for diagnosing DKD and a potential target for treatment.