The First Clinical Medical College, Zhejiang Chinese Medical University, Hangzhou, China.
Department of Cardiology, the First Affiliated Hospital of Zhejiang Chinese Medical University, Zhejiang Provincial Hospital of Chinese Medicine, Hangzhou, China.
Front Immunol. 2024 Feb 19;15:1353111. doi: 10.3389/fimmu.2024.1353111. eCollection 2024.
Myocardial infarction (MI) caused by severe coronary artery disease has high incidence and mortality rates, making its prevention and treatment a central and challenging aspect of clinical work for cardiovascular practitioners. Recently, researchers have turned their attention to a novel mechanism of cell death caused by Cu, cuproptosis.
This study integrated data from three MI-related bulk datasets downloaded from the Gene Expression Omnibus (GEO) database, and identified 16 differentially expressed genes (DEGs) related to cuproptosis by taking intersection of the 6378 DEGs obtained by differential analysis with 49 cuproptosis-related genes. Four hub genes, Dbt, Dlat, Ube2d1 and Ube2d3, were screened out through random forest analysis and Lasso analysis. In the disease group, Dbt, Dlat, and Ube2d1 showed low expression, while Ube2d3 exhibited high expression.
Focusing on Ube2d3 for subsequent functional studies, we confirmed its high expression in the MI group through qRT-PCR and Western Blot detection after successful construction of a MI mouse model by left anterior descending (LAD) coronary artery ligation, and further clarified the correlation of cuproptosis with MI development by detecting the levels of cuproptosis-related proteins. Moreover, through experiments, Ube2d3 was confirmed to be highly expressed in oxygen-glucose deprivation (OGD)-treated cardiomyocytes AC16. In order to further clarify the role of Ube2d3, we knocked down Ube2d3 expression in OGD-treated AC16 cells, and confirmed Ube2d3's promoting role in the hypoxia damage of AC16 cells by inducing cuproptosis, as evidenced by the detection of MTT, TUNEL, LDH release and cuproptosis-related proteins.
In summary, our findings indicate that Ube2d3 regulates cuproptosis to affect the progression of MI.
由严重冠状动脉疾病引起的心肌梗死（MI）发病率和死亡率较高，使其成为心血管医生临床工作的重点和难点。最近，研究人员将注意力转向一种由 Cu 引起的新型细胞死亡机制，即铜死亡。
本研究整合了从基因表达综合数据库（GEO）下载的三个与 MI 相关的批量数据集的数据，通过取差异分析得到的 6378 个差异表达基因（DEGs）与 49 个铜死亡相关基因的交集，鉴定出 16 个与铜死亡相关的 DEGs。通过随机森林分析和 Lasso 分析筛选出 4 个枢纽基因：Dbt、Dlat、Ube2d1 和 Ube2d3。在疾病组中，Dbt、Dlat 和 Ube2d1 表达较低，而 Ube2d3 表达较高。
针对 Ube2d3 进行后续功能研究，通过左前降支（LAD）冠状动脉结扎成功构建 MI 小鼠模型后，通过 qRT-PCR 和 Western Blot 检测，我们确认了 Ube2d3 在 MI 组中的高表达，并通过检测铜死亡相关蛋白，进一步阐明了铜死亡与 MI 发生的相关性。此外，通过实验证实 Ube2d3 在氧葡萄糖剥夺（OGD）处理的心肌细胞 AC16 中高表达。为了进一步阐明 Ube2d3 的作用，我们在 OGD 处理的 AC16 细胞中敲低 Ube2d3 的表达，通过诱导铜死亡，证实 Ube2d3 通过促进铜死亡对 AC16 细胞的缺氧损伤发挥作用，这可以通过 MTT、TUNEL、LDH 释放和铜死亡相关蛋白的检测来证明。
综上所述，我们的研究结果表明，Ube2d3 通过调节铜死亡来影响 MI 的进展。
