Department of Pediatrics, Shuguang Hospital Affiliated to Shanghai Traditional Chinese Medical University, No. 528 Zhangheng Road, Pudong New Area, Shanghai, 201203, China.
Department of Clinical Laboratory, Shuguang Hospital Affiliated to Shanghai Traditional Chinese Medical University, Shanghai, 201203, China.
Cell Tissue Res. 2019 Oct;378(1):67-80. doi: 10.1007/s00441-019-03026-6. Epub 2019 May 18.
The aim of this study is to investigate the potential roles of miR-943-3p and its target gene secreted frizzled-related proteins4 (SFRP4) in allergic asthma and elucidate its underlying mechanism, which may prompt a new clue about developing novel treatments of this disease. An allergic asthma mouse model was generated by challenging with ovalbumin (OVA); lung pathological features of mice were viewed using H&E staining; thickness of subepithelial fibrosis and smooth muscle was measured using Masson's trichrome staining. Inflammatory cells from bronchoalveolar lavage fluid (BALF) were counted based on Diff-Quik staining and morphometric analysis. Expressions of miR-943-3p, SFRP4 and Wnt signal pathway-associated proteins were detected using RT-PCR or immunoblotting, respectively. SFRP4 was downregulated in the bronchial biopsies of allergic asthma patients and represented a unique intersection between differentially expressed genes (DEGs) and genes in the Wnt signal pathway. Both miR-943-3p upregulation and SFRP4 downregulation were detected in allergic asthma patients and OVA-induced mice. Besides, OVA-induced mice possessed more inflammatory cells in BALF including macrophage (mac), eosinophil (eos), lymphocyte (lym) and neutrophil (neu), higher expression of collagen, β-catenin and c-Myc as well as thicker subepithelial fibrosis and smooth muscle in lung than control mice. In vivo delivery of miR-943-3p agomir worsened these symptoms, while both miR-943-3p antagomir and Ad-SFRP4 administration effectively alleviated this disease. Taken together, miR-943-3p accelerated the progression of airway inflammation and remodeling in allergic asthma via suppressing the activity of SFRP4 through Wnt signaling pathway in asthma patients and OVA-induced mice.
本研究旨在探讨 miR-943-3p 及其靶基因分泌型卷曲相关蛋白 4（SFRP4）在过敏性哮喘中的潜在作用，并阐明其潜在机制，这可能为开发这种疾病的新疗法提供新线索。通过卵清蛋白（OVA）挑战构建过敏性哮喘小鼠模型；使用 H&E 染色观察小鼠肺部病理特征；使用 Masson 三色染色测量上皮下纤维化和平滑肌的厚度。根据 Diff-Quik 染色和形态计量分析计数支气管肺泡灌洗液（BALF）中的炎性细胞。使用 RT-PCR 或免疫印迹分别检测 miR-943-3p、SFRP4 和 Wnt 信号通路相关蛋白的表达。在过敏性哮喘患者的支气管活检组织中发现 SFRP4 下调，并且它代表了差异表达基因（DEGs）和 Wnt 信号通路基因之间的独特交点。在过敏性哮喘患者和 OVA 诱导的小鼠中均检测到 miR-943-3p 上调和 SFRP4 下调。此外，OVA 诱导的小鼠在 BALF 中具有更多的炎性细胞，包括巨噬细胞（mac）、嗜酸性粒细胞（eos）、淋巴细胞（lym）和中性粒细胞（neu），肺中胶原、β-连环蛋白和 c-Myc 的表达更高，上皮下纤维化和平滑肌更厚，与对照小鼠相比。体内给予 miR-943-3p 激动剂会加重这些症状，而给予 miR-943-3p 拮抗剂和 Ad-SFRP4 治疗均可有效缓解该疾病。综上所述，miR-943-3p 通过抑制 Wnt 信号通路中 SFRP4 的活性，加速了哮喘患者和 OVA 诱导的小鼠气道炎症和重塑的进展。
