摘要目的:探讨miR-15a-5p调控Wnt信号通路对PQ致肺纤维化的影响及分子机制。方法:构建PQ诱导的16HBE细胞模型，采用高通量miRNA芯片技术和RT-qPCR筛选表达差异明显的miR-15a-5p进行实验。实验分组为：NC组（对照组）：无特殊处理；PQ组：50 μmol/L PQ处理细胞72 h；miR-15a-5p组：转染miR-15a-5p过表达慢病毒的16HBE稳转株；miR-15a-5p+PQ组：50 μmol/L PQ处理稳转株细胞72 h。RT-qPCR和Western blot检测Wnt通路相关基因Wnt3α和β-catenin、成纤维细胞标记基因Collagen I、Vimentin和α SMA，上皮细胞标记基因Occludin和CK18表达情况。构建PQ诱导的肺纤维化小鼠模型，采用Western blot、HE染色和免疫组织化学检测蛋白表达及肺组织损伤情况。数据以均数±标准差（ ± s）表示，采用独立样本 t检验分析两组间数据。 结果:细胞损伤模型中，Wnt3α、β-catenin、成纤维细胞标记基因Collagen I、Vimentin和α SMA表达显著上调（ P<0.05），上皮细胞标记基因Occludin和CK18显著下调（ P<0.05），过表达miR-15a-5p可靶向抑制Wnt3α表达并缓解PQ诱导的EMT进程。动物模型中，Wnt3α、β-catenin、成纤维细胞标记基因Collagen I、Vimentin和α SMA蛋白水平显著升高（ P<0.01），肺组织结构紊乱并发生纤维化，过表达miR-15a-5p可抑制Wnt3α蛋白表达水平（ P<0.05）且改善肺组织损伤。 结论:miR-15a-5p可通过调控Wnt3α/β-catenin信号通路改善PQ导致的肺损伤，从而抑制肺纤维化的发生发展。

abstractsObjective:To investigate the effect and molecular mechanism of miR-15a-5p regulation Wnt signaling pathway in PQ-induced pulmonary fibrosis.Methods:The PQ-induced 16HBE cell model was constructed, high-throughput miRNA chip and RT-qPCR were used to screen for miR-15a-5p with significant differences. The experimental groups were as follows: NC group (normal control);no special treatment; PQ group: 50 μmol/L PQ treated cells for 72 h; miR-15a-5p group: 16HBE stable cell lines transfected with miR-15a-5p overexpressing lentivirus; miR-15a-5p+PQ group: Stable cell lines were treated with 50 μmol/L PQ for 72 h. The expression of Wnt pathway-related genes Wnt3α and β-catenin, fibroblast marker genes Collagen I, Vimentin and α SMA, epithelial marker genes Occludin and CK18 were detected by RT-qPCR and Western blot. The mice model of PQ-induced pulmonary fibrosis was constructed, and the protein expression and lung tissue injury were detected by Western blot, HE staining and immunohistochemistry. Data were expressed as mean ± standard deviation, and independent sample t-test was used to analyze the data between the two groups. Results:The expressions of Wnt3α, β-catenin, fibroblast marker genes Collagen I, Vimentin and α SMA significantly up-regulated in cell injury models ( P<0.05), the epithelial cell marker genes Occludin and CK18 significantly down-regulated ( P<0.05), overexpression of miR-15a-5p could inhibit the expression of Wnt3α and alleviated the EMT induced by PQ. In animal models, Wnt3α, β-catenin, fibroblast marker genes Collagen I, Vimentin and α SMA significantly increased ( P<0.01), the structure of lung tissue was disordered and fibrosis occurred, overexpression of miR-15a-5p inhibited the expression of Wnt3α protein ( P<0.05) and ameliorated lung tissue injury. Conclusions:miR-15a-5p ameliorates PQ-induced lung injury by modulating the Wnt3α/β-catenin signaling pathway, thereby inhibiting the development of pulmonary fibrosis.