Department of Respiratory and Critical Care Medicine, Henan Provincial People's Hospital, People's Hospital of Zhengzhou University, School of Clinical Medicine, Henan University, Zhengzhou City, China.
Chem Biol Drug Des. 2024 Mar;103(3):e14508. doi: 10.1111/cbdd.14508.
Pulmonary Fibrosis (PF) is a fatal lung disease with complicated pathogenesis. Astragaloside IV (ASV) has been discovered to alleviate PF progression, and the potential molecular mechanism of ASV in the development of PF need to be further clarified. Bleomycin (BLM) was used to construct PF in vivo model. Expression levels of circ_0008898, miR-211-5p, high mobility group protein B1 (HMGB1), alpha smooth muscle Actin (α-SMA) and Collagen I were examined by Quantitative real time polymerase chain reaction (qRT-PCR) and western blot. Cell survival was analyzed using Cell Counting Kit-8 (CCK-8) and EdU (5-ethynyl-2'-deoxyuridine) assay. The invasion abilities were investigated by transwell assay. The levels of inflammatory factors were tested via using Enzyme-linked immunosorbent assay (ELISA). The relationship between circ_0008898 or HMGB1 and miR-211-5p was identified by dual-luciferase reporter assay. The results showed that ASV attenuated BLM-induced pulmonary fibrosis in vivo. In vitro study, ASV alleviated TGF-β1-induced fibrogenesis in HFL1 cells. Circ_0008898 was increased in TGF-β1-induced HFL1 cells. ASV-induced impacts were abrogated by circ_0008898 overexpression in TGF-β1-induced HFL1 cells. Mechanistically, circ_0008898 competitively bound to miR-211-5p to increase the expression of its target HMGB1. MiR-211-5p deficiency rescued ASV-mediated effects in TGF-β1-induced HFL1 cells. In addition, HMGB1 overexpression partially overturned circ_0008898 interference-induced impacts in HFL1 cells upon TGF-β1 treatment. In conclusion, our work manifested that ASV hindered PF process by mediating the circ_0008898/miR-211-5p/HMGB1 network.
肺纤维化（PF）是一种具有复杂发病机制的致命肺部疾病。已发现黄芪甲苷（ASV）可减轻 PF 的进展，需要进一步阐明 ASV 在 PF 发展中的潜在分子机制。博来霉素（BLM）用于构建体内 PF 模型。通过定量实时聚合酶链反应（qRT-PCR）和蛋白质印迹法检测 circ_0008898、miR-211-5p、高迁移率族蛋白 B1（HMGB1）、α平滑肌肌动蛋白（α-SMA）和 I 型胶原的表达水平。使用细胞计数试剂盒-8（CCK-8）和 5-乙炔基-2'-脱氧尿苷（EdU）测定法分析细胞存活率。通过 Transwell 测定法研究细胞侵袭能力。通过酶联免疫吸附测定法（ELISA）测试炎症因子水平。通过双荧光素酶报告基因检测法鉴定 circ_0008898 或 HMGB1 与 miR-211-5p 之间的关系。结果表明，ASV 可减轻 BLM 诱导的体内肺纤维化。在体外研究中，ASV 减轻 TGF-β1 诱导的 HFL1 细胞纤维化。TGF-β1 诱导的 HFL1 细胞中 circ_0008898 增加。在 TGF-β1 诱导的 HFL1 细胞中过表达 circ_0008898 可消除 ASV 诱导的作用。机制上，circ_0008898 竞争性结合 miR-211-5p 以增加其靶标 HMGB1 的表达。miR-211-5p 缺陷挽救了 TGF-β1 诱导的 HFL1 细胞中 ASV 介导的作用。此外，HMGB1 过表达部分逆转了 TGF-β1 处理后 HFL1 细胞中转录本 0008898 干扰诱导的影响。总之，我们的工作表明，ASV 通过介导 circ_0008898/miR-211-5p/HMGB1 网络来抑制 PF 进程。
