Fang Xiansong, Wen Xiaoyun, Zhou Liang, Jiang Yingjie, Wang Liefeng
Department of Blood Transfusion, First Affiliated Hospital of Gannan Medical University, Ganzhou, China.
Department of Biochemistry and Molecular Biology, Gannan College of Medicine, China Medical University, Shenyang, China.
Cytojournal. 2025 Apr 1;22:37. doi: 10.25259/Cytojournal_223_2024. eCollection 2025.
Idiopathic pulmonary fibrosis (PF) is a chronic and life-threatening lung disease. This study aimed to investigate the role of zinc finger and BTB domain containing 16 (Zbtb16), a transcription factor, in the progression of PF by analyzing its expression and regulatory effects in mouse and cell models.
The gene expression profiles in bleomycin-induced (BL-I) PF lung tissues of mice were analyzed using the gene expression omnibus database. The mouse model of BL-I PF and cell model of transforming growth factor-β1 (TGF-β1)-induced mice lung epithelial cell (LEC) fibrosis was constructed. Zbtb16 expression was evaluated by reverse transcription quantitative polymerase chain reaction, Western blot, or immunohistochemistry. Tissue sections were assessed by hematoxylin and eosin, Masson, and terminal deoxynucleotidyl transferase dUTP nick-end labeling staining. The levels of protein, inflammation factors, and albumin were measured through Western blot or enzyme-linked immunosorbent assay.
Bioinformatics analysis found that Zbtb16 was the highest differentially expressed marker in BL-I PF mice. Zbtb16 was highly expressed in the mice and cell model. Zbtb16 silencing could reduce lung tissues' collagen deposition, pulmonary edema, and pulmonary apoptotic cells; improve vascular permeability; and decrease fibrosis markers and inflammation factors expressed in model mice. Zbtb16 silencing could reduce fibrosis markers and inflammation factor levels in the cell model ( < 0.05). Kyoto encyclopedia of genes and genomes and gene set enrichment analyses suggested that Zbtb16 might regulate BL-I PF in mice through the phosphoinositide 3-kinases (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway (PAmT-P). Co-immunoprecipitation showed the combination of AKT and Zbtb16. PAmT-P in the mice model and cell model was visibly activated ( < 0.05), and Zbtb16 silencing could inhibit it ( < 0.05). Moreover, the rescue experiments showed that the AKT activator SC79 could reverse the effect of TGF-β1 + small interfere RNA-Zbtb16 on LECs.
This study identified Zbtb16 as a key regulator of PF progression, mediating its effects through the PAmT-P. Zbtb16 silencing alleviated fibrosis and inflammation and , providing a promising target for therapeutic intervention in PF.
特发性肺纤维化（PF）是一种慢性且危及生命的肺部疾病。本研究旨在通过分析转录因子锌指和BTB结构域包含蛋白16（Zbtb16）在小鼠和细胞模型中的表达及调控作用，探讨其在PF进展中的作用。
利用基因表达综合数据库分析博来霉素诱导（BL-I）的PF小鼠肺组织中的基因表达谱。构建BL-I PF小鼠模型和转化生长因子-β1（TGF-β1）诱导的小鼠肺上皮细胞（LEC）纤维化细胞模型。通过逆转录定量聚合酶链反应、蛋白质免疫印迹或免疫组织化学评估Zbtb16表达。用苏木精-伊红、Masson和末端脱氧核苷酸转移酶dUTP缺口末端标记染色评估组织切片。通过蛋白质免疫印迹或酶联免疫吸附测定法测量蛋白质、炎症因子和白蛋白水平。
生物信息学分析发现Zbtb16是BL-I PF小鼠中差异表达最高的标志物。Zbtb16在小鼠和细胞模型中高表达。Zbtb16沉默可减少肺组织中的胶原沉积、肺水肿和肺凋亡细胞；改善血管通透性；并降低模型小鼠中表达的纤维化标志物和炎症因子。Zbtb16沉默可降低细胞模型中的纤维化标志物和炎症因子水平（<0.05）。京都基因与基因组百科全书和基因集富集分析表明，Zbtb16可能通过磷酸肌醇3-激酶（PI3K）/蛋白激酶B（AKT）/雷帕霉素哺乳动物靶蛋白（mTOR）途径（PAmT-P）调控小鼠中的BL-I PF。免疫共沉淀显示AKT与Zbtb16结合。小鼠模型和细胞模型中的PAmT-P明显被激活（<0.05），Zbtb16沉默可抑制它（<0.05）。此外，拯救实验表明AKT激活剂SC79可逆转TGF-β1 + 小干扰RNA-Zbtb16对LEC的作用。
本研究确定Zbtb16是PF进展的关键调节因子，通过PAmT-P介导其作用。Zbtb16沉默减轻了纤维化和炎症，为PF的治疗干预提供了一个有前景的靶点。
