Lian Ningfang, Zhang Qiaoxian, Chen Jia, Chen Mengxue, Huang Jiefeng, Lin Qichang
Department of Pulmonary and Critical Care Medicine, The First Affiliated Hospital of Fujian Medical University, Institute of Respiratory Disease, Fujian Medical University, Fuzhou, China.
Front Physiol. 2021 Sep 29;12:751206. doi: 10.3389/fphys.2021.751206. eCollection 2021.
Cigarette smoking is a major risk factor for bronchoalveolar epithelial cell (BAEC) injury. Understanding the relevant pathogenesis is important for the treatment of cigarette smoke-related chronic airway diseases such as chronic obstructive pulmonary disease. In this study, BAECs were cultured in 5% cigarette smoke extract (CSE) or regular culture medium for 24 h. Differentially expressed genes (DEGs) were detected by next-generation RNA sequencing (RNA-seq) and validated by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Bioinformatic analysis was performed on DEGs. Co-treated BAECs with 5% CSE and the ferroptosis inhibitor, ferrostatin-1 was applied to observe the role of ferroptosis. In the CSE group, 210 upregulated genes and 159 downregulated genes were identified compared with the control group. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed that the DEGs were related to oxidative stress and ferroptosis. Ferroptosis-related genes were further verified by qRT-PCR. The mRNA level of GPX4 decreased; the mRNA levels of ACSL4, FTH1 and SLC7A11 increased ( < 0.05). Pretreatment with the ferroptosis inhibitor ferrostatin-1 mitigated CSE-induced ROS accumulation and inflammatory mediator expression in BAECs ( < 0.05). CSE treatment altered ferroptosis-related gene expression patterns in cultured BAECs. Inhibition of ferroptosis reduced the inflammatory response of CSE-treated BAECs. These data provide a better understanding of the underlying molecular mechanisms of CSE-related lung injury.
吸烟是支气管肺泡上皮细胞（BAEC）损伤的主要危险因素。了解相关发病机制对于治疗与香烟烟雾相关的慢性气道疾病（如慢性阻塞性肺疾病）至关重要。在本研究中，将BAECs在5%香烟烟雾提取物（CSE）或常规培养基中培养24小时。通过下一代RNA测序（RNA-seq）检测差异表达基因（DEGs），并通过定量逆转录聚合酶链反应（qRT-PCR）进行验证。对DEGs进行生物信息学分析。用5% CSE和铁死亡抑制剂铁抑素-1共同处理BAECs，以观察铁死亡的作用。与对照组相比，在CSE组中鉴定出210个上调基因和159个下调基因。基因本体论和京都基因与基因组百科全书（KEGG）分析表明，DEGs与氧化应激和铁死亡有关。通过qRT-PCR进一步验证铁死亡相关基因。GPX4的mRNA水平降低；ACSL4、FTH1和SLC7A11的mRNA水平升高（<0.05）。用铁死亡抑制剂铁抑素-1预处理可减轻CSE诱导的BAECs中ROS积累和炎症介质表达（<0.05）。CSE处理改变了培养的BAECs中铁死亡相关基因的表达模式。抑制铁死亡可降低CSE处理的BAECs的炎症反应。这些数据有助于更好地理解CSE相关肺损伤的潜在分子机制。
