Duan Xiaomei, Hu Tingting, Xu Lijuan, Li Zheng, Jing Jing, Xu Dan, Ding Jianbing, Li Fengsen, Jiang Min, Wang Jing
Department of Xinjiang Laboratory of Respiratory Disease Research, Traditional Chinese Medicine Hospital Affiliated to Xinjiang Medical University, Urumqi, Xinjiang, China.
Department of Xinjiang Clinical Research Center for Respiratory Diseases, Traditional Chinese Medicine Hospital Affiliated to Xinjiang Medical University, Urumqi, Xinjiang, China.
Immun Inflamm Dis. 2024 Dec;12(12):e70104. doi: 10.1002/iid3.70104.
Chronic obstructive pulmonary disease (COPD), a prevalent respiratory condition, is characterized by long-term airway inflammation, which can lead to airway remodeling and persistent airflow restriction. Exposure to cigarette smoke is known as a major contributor to COPD development. Research has confirmed that ferroptosis and m6A modification are closely related to various inflammatory-related diseases. However, the correlation between m6A methylation and ferroptosis in COPD has not been confirmed. In this study, combined with bioinformatics analysis and molecular biology methods we investigated how m6A methylation was correlated to ferroptosis-associated genes (SLC7A11 and NQO-1) in cigarette smoke induced 16HBES cells.
Two microarray datasets (GSE30063 and GSE64614) were combined to identify differentially expressed genes (DEGs) through the application of bioinformatics techniques. A cigarette smoke (CS)-induced 16HBE cells model was established. The ROS, GSH, MDA, and total iron content were detected by relevant detection kits. The expression levels associated with ferroptosis and m6A methylation modification-related genes were determined via reverse transcription-quantitative polymerase chain reaction and western blot.
Overall, 529 DEGs were identified in the above two databases. For COPD patients, significant changes were observed in FAGs (GCLC, NQO-1, SLC7A11) and m6A methylation-related genes (FTO). A negative correlation was also noted between the expression level of genes linked to ferroptosis (SLC7A11 and NQO-1) and that of the m6A methylation gene (FTO). The in vitro experiments results indicate that SLC7A11 and NQO-1 were significantly downregulated, and FTO were significantly upregulated. In addition, cigarette smoke stimulation increased the levels of MDA, LPO, and ROS, while reducing the content of GSH and total iron content in 16HBE cells.
Our findings explored the relationship between ferroptosis and m6A methylation in COPD, and screened out SLC7A11, NQO-1 and FTO may be critical in the pathogenesis of COPD.
慢性阻塞性肺疾病（COPD）是一种常见的呼吸道疾病，其特征为长期气道炎症，可导致气道重塑和持续性气流受限。接触香烟烟雾是COPD发病的主要促成因素。研究证实，铁死亡和m6A修饰与多种炎症相关疾病密切相关。然而，COPD中m6A甲基化与铁死亡之间的相关性尚未得到证实。在本研究中，我们结合生物信息学分析和分子生物学方法，研究了香烟烟雾诱导的16HBES细胞中m6A甲基化与铁死亡相关基因（SLC7A11和NQO-1）的相关性。
通过应用生物信息学技术，合并两个微阵列数据集（GSE30063和GSE64614）以鉴定差异表达基因（DEG）。建立香烟烟雾（CS）诱导的16HBE细胞模型。使用相关检测试剂盒检测活性氧（ROS）、谷胱甘肽（GSH）、丙二醛（MDA）和总铁含量。通过逆转录定量聚合酶链反应和蛋白质印迹法测定与铁死亡和m6A甲基化修饰相关基因的表达水平。
总体而言，在上述两个数据库中鉴定出529个DEG。对于COPD患者，在铁死亡相关基因（GCLC、NQO-1、SLC7A11）和m6A甲基化相关基因（FTO）中观察到显著变化。还注意到与铁死亡相关的基因（SLC7A11和NQO-1）的表达水平与m6A甲基化基因（FTO）的表达水平呈负相关。体外实验结果表明，SLC7A11和NQO-1显著下调，而FTO显著上调。此外，香烟烟雾刺激增加了16HBE细胞中MDA、脂质过氧化（LPO）和ROS的水平，同时降低了GSH含量和总铁含量。
我们的研究结果探讨了COPD中铁死亡与m6A甲基化之间的关系，并筛选出SLC7A11、NQO-1和FTO可能在COPD发病机制中起关键作用。
