背景:这项研究的目的是鉴定椎间盘退变(IDD)过程中与纤维环(AF)中铁凋亡相关的差异表达基因(DEGs)。
方法:我们分析了从GSE70362和GSE147383数据集获得的退化和正常AF的基因数据。进行了分析以确定DEG的功能意义,随后创建了一个网络,说明蛋白质之间的相互作用。我们进一步分析了DEGs的免疫浸润,并使用LASSO回归分析确定了中心DEGs。最后,我们鉴定了枢纽铁凋亡相关的DEGs(FRDEGs),并使用实时定量聚合酶链反应(RT-qPCR)验证了它们的表达水平,蛋白质印迹,免疫组织化学染色(IHC),和免疫荧光(IF)。
结果:通过分析GSE70362和GSE147383数据集,我们确定了118个DEG。在退行性房颤组中,我们观察到静息记忆CD4+T细胞的免疫浸润明显增加。LASSO回归分析显示有9个集线器DEG。接收器工作特性(ROC)曲线的构建产生0.762的曲线下面积(AUC)值。此外,我们发现MGST1是一个与铁凋亡相关的hub基因。我们对免疫浸润的检查表明,MGST1主要影响不同免疫细胞表达组中的巨噬细胞M0。最后,我们的观察显示MGST1在变性纤维环组织中的表达显著上调.
结论:我们的研究结果表明,退化性房颤中MGST1水平激增,可能在IDD的恶化中起着至关重要的作用。这些发现为进一步探索IDD的病理机制奠定了基础,并为干预提供了潜在的药物靶标。
BACKGROUND: The objective of this research was to identify differentially expressed genes (DEGs) related to ferroptosis in the annulus fibrosus (AF) during intervertebral disc degeneration (IDD).
METHODS: We analyzed gene data from degenerated and normal AF obtained from the GSE70362 and GSE147383 datasets. An analysis to determine the functional significance of the DEGs was conducted, followed by the creation of a network illustrating the interactions between proteins. We further analyzed the immune infiltration of the DEGs and determined the hub DEGs using LASSO regression analysis. Finally, we identified the hub ferroptosis-related DEGs (FRDEGs) and verified their expression levels using Real-time quantitative polymerase chain reaction (RT-qPCR), Western blot, Immunohistochemical Staining (IHC), and Immunofluorescence (IF).
RESULTS: By analyzing the GSE70362 and GSE147383 datasets, we identified 118 DEGs. In degenerative AF groups, we observed a significant increase in immune infiltration of resting memory CD4+ T cells. LASSO regression analysis revealed 9 hub DEGs. The construction of a Receiver Operating Characteristic (ROC) curve yielded an Area Under the Curve (AUC) value of 0.762. Furthermore, we found that MGST1 is a hub gene related to ferroptosis. Our examination of immune infiltration indicated that MGST1 primarily influences macrophage M0 in different immune cell expression groups. Finally, our observations revealed a marked upregulation of MGST1 expression in the degenerated annulus fibrosus tissue.
CONCLUSIONS: Our findings indicate an upsurge in MGST1 levels within degenerative AF, potentially playing a crucial role in the exacerbation of IDD. These findings provide a foundation for further exploration of the pathological mechanisms underlying IDD and offer potential drug targets for intervention.