Objective Acute respiratory distress syndrome(ARDS)patients currently have relatively high mortality,which is associated with early lung fibrosis.This study aimed to investigate whether miR-17 suppression could allevi...Objective Acute respiratory distress syndrome(ARDS)patients currently have relatively high mortality,which is associated with early lung fibrosis.This study aimed to investigate whether miR-17 suppression could alleviate ARDS-associated lung fibrosis by regulating Mfn2.Methods A mouse model of ARDS-related lung fibrosis was constructed via intratracheal instillation of bleomycin.The expression level of miR-17 in lung tissues was detected via quantitative real time polymerase chain reaction(qRT-PCR).In the ARDS mouse model of lung fibrosis,the mitigating effects of miR-17 interference were evaluated via tail vein injection of the miR negative control or the miR-17 antagomir.The pathological changes in the lung tissue were examined via HE staining and Masson’s trichrome staining,and the underlying molecular mechanism was investigated via ELISA,qRT-PCR and Western blotting.Results Bleomycin-induced pulmonary fibrosis significantly increased collagen deposition and the levels of hydroxyproline(HYP)and miR-17.Interfering with miR-17 significantly reduced the levels of HYP and miR-17 and upregulated the expression of Mfn2.The intravenous injection of the miR-17 antagomir alleviated lung inflammation and reduced collagen deposition.In addition,interference with miR-17 could upregulate LC3B expression,downregulate p62 expression,and improve mitochondrial structure.Conclusion Interfering with miR-17 can improve pulmonary fibrosis in mice by promoting mitochondrial autophagy via Mfn2.展开更多
基金supported by grants from the Natural Science Foundation of Hubei Province(No.2022CFB423,2023AFB1055)Hubei Province Health and Family Planning Scientific Research Project(No.WJ2023M030).
文摘Objective Acute respiratory distress syndrome(ARDS)patients currently have relatively high mortality,which is associated with early lung fibrosis.This study aimed to investigate whether miR-17 suppression could alleviate ARDS-associated lung fibrosis by regulating Mfn2.Methods A mouse model of ARDS-related lung fibrosis was constructed via intratracheal instillation of bleomycin.The expression level of miR-17 in lung tissues was detected via quantitative real time polymerase chain reaction(qRT-PCR).In the ARDS mouse model of lung fibrosis,the mitigating effects of miR-17 interference were evaluated via tail vein injection of the miR negative control or the miR-17 antagomir.The pathological changes in the lung tissue were examined via HE staining and Masson’s trichrome staining,and the underlying molecular mechanism was investigated via ELISA,qRT-PCR and Western blotting.Results Bleomycin-induced pulmonary fibrosis significantly increased collagen deposition and the levels of hydroxyproline(HYP)and miR-17.Interfering with miR-17 significantly reduced the levels of HYP and miR-17 and upregulated the expression of Mfn2.The intravenous injection of the miR-17 antagomir alleviated lung inflammation and reduced collagen deposition.In addition,interference with miR-17 could upregulate LC3B expression,downregulate p62 expression,and improve mitochondrial structure.Conclusion Interfering with miR-17 can improve pulmonary fibrosis in mice by promoting mitochondrial autophagy via Mfn2.
文摘目的对人类miR-17-5p(hsa-miR-17-5p)靶基因预测和富集分析,并结合基因的差异表达寻找多发性硬化症的生物学标志物。方法利用生物信息学方法,预测hsa-miR-17-5p靶基因,对预测结果进行基因本体数据库(Gene Ontology,GO)、京都基因和基因组百科全书(Kyoto Encyclopedia of Genes and Genome,KEGG)富集分析,并构建蛋白质互作网络图(Protein-Protein Interaction Networks,PPI)。再结合多发性硬化症患者和健康对照组的差异表达基因,进一步筛选出hsa-miR-17-5p对多发性硬化症影响的关键基因。最后,利用RNA22 v2验证关键基因是否存在结合位点。结果不同网站预测后,取交集获取376个公共靶基因,这些靶基因有复杂的相互作用,主要存在于胞浆与核质,参与多种转录调控和蛋白质磷酸化等生物学过程,同时也与癌症、病毒感染和免疫系统等信号通路相关;靶基因和多发性硬化症患者的相对下调基因有三个共同基因PDLIM5、KCNB1、BTBD7。进一步验证上述关键基因结合位点,发现仅有PDLIM5存在P值小于0.05的结合位点。结论hsa-miR-17-5p靶基因参与多种生物学过程,而且PDLIM5、KCNB1、BTBD7基因很可能是hsa-miR-17-5p对多发性硬化症产生影响的重要桥梁,其中PDLIM5尤为重要。
