摘要： 目的 探讨miR-23b-5p靶向下调纤维连接蛋白1（FN1）对支气管哮喘（简称哮喘）气道重塑的影响。方法 将BALB/c小鼠随机分为正常对照组和哮喘组，每组6只。Masson染色观察气道壁周围胶原沉积水平。ELISA测定白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）、转化生长因子-β1（TGF-β1）、血管内皮生长因子（VEGF）水平。实时荧光定量PCR（qRT-PCR）检测肺组织中miR-23b-5p表达水平。免疫组织化学染色、Western blotting检测FN1、α-平滑肌肌动蛋白（α-SMA）在肺组织中的表达。TargetScan网站预测miR-23b-5p靶基因，双荧光素酶报告实验验证。转染miR-23b-5p模拟物和抑制剂于人支气管平滑肌细胞（HBSMC）后，Western blotting检测FN1表达水平。结果 与正常对照组相比，哮喘组小鼠气道上皮下胶原沉积，IL-6、TNF-α、TGF-β1、VEGF表达显著增高（P<0.001），miR-23b-5p表达显著降低（P<0.05）。免疫组织化学染色、Western blotting结果显示，FN1、α-SMA在哮喘组小鼠肺组织中的表达显著升高（P<0.05）。FN1受miR-23b-5p靶向调控，并且与miR-NC组相比，转染miR-23b-5p模拟物后FN1表达水平显著下调（P<0.05），而转染miR-23b-5p抑制剂后FN1表达水平显著升高（P<0.01）。结论 miR-23b-5p可通过靶向下调FN1，缓解哮喘气道重塑。

miR-23b-5p通过靶向下调FN1缓解支气管哮喘气道重塑

白诗瑶, 包慧静, 马琳, 麻雪晴, 李丹玲, 苏新明

中国医科大学附属第一医院呼吸与危重症医学科, 沈阳 110001

收稿日期:2022-11-22出版日期:2023-06-30发布日期:2023-05-31
通讯作者:苏新明E-mail:xinming_s@163.com
作者简介:白诗瑶（1995-），女，医师，硕士
基金资助:国家自然科学基金（82170038）


关键词: 支气管哮喘, 气道重塑, miR-23b-5p, 纤维连接蛋白1
Abstract: Objective To investigate the effects of miR-23b-5p on airway remodeling in bronchial asthma through targeted downregulation of fibronectin 1 (FN1). Methods BALB/c mice were randomly divided into a control or an asthma group (n=6 mice per group). Collagen deposition in the airway walls were detected using Masson staining. The levels of interleukin-6 (IL-6),tumor necrosis factor-α (TNF-α),transforming growth factor-β1 (TGF-β1),and vascular endothelial growth factor (VEGF) were determined using ELISA. Realtime fluorescence quantitative PCR (qRT-PCR) was performed to detect miR-23b-5p expression levels in lung tissues. Immunohistochemical staining and Western blotting were performed to detect the expression of FN1 and α-smooth muscle actin (α-SMA) in lung tissues. miR-23b-5p target genes were predicted using the TargetScan website,and validated using dual luciferase reporter assay. Western blotting was performed to detect the FN1 expression levels after transfection of miR-23b-5p mimic and inhibitor in human bronchial smooth muscle cells. Results Compared with the control group,mice in the asthma group had subepithelial collagen deposition in their airway, significantly higher levels of IL-6,TNF-α,TGF-β1,and VEGF (P < 0.001),and significantly lower levels of miR-23b-5p (P < 0.05). Immunohistochemical staining and Western blotting showed that the expression of FN1 and α-SMA significantly increased in the asthma group (P < 0.05). FN1 was directly regulated by miR-23b-5p. Compared with the miR-NC group,FN1 expression levels were significantly down-regulated after transfection with miR-23b-5p mimic (P < 0.05) and significantly increased after transfection with miR-23b-5p inhibitor (P < 0.01). Conclusion miR-23b-5p relieves airway remodeling in bronchial asthma through the downregulation of FN1.
Key words: bronchial asthma, airway remodeling, miR-23b-5p, fibronectin 1
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