中国全科医学2025，Vol.28Issue(25)：3151-3160,10.DOI:10.12114/j.issn.1007-9572.2023.0766
Jagged1/Notch1信号通路对特发性肺纤维化过程中内皮间质转化的影响研究
The Impact of the Jagged1/Notch1 Signalling Pathway on Endothelial-mesenchymal Transition in Idiopathic Pulmonary Fibrosis
摘要
Abstract
Background Idiopathic pulmonary fibrosis(IPF)is a complex global disease with a pathological mechanism that remains poorly understood and for which there is currently no effective treatment.Endothelial-mesenchymal transition(EndMT),identified as a pathological basis for various chronic vascular diseases,has recently been recognised as a significant risk factor for IPF.However,the related mechanisms have yet to be clarified.Objective To observe the effects of the Jagged1/Notch1 signalling pathway on EndMT to elucidate the related mechanisms during IPF.Methods From March to June 2021,human pulmonary arterial endothelial cells(HPAEC)were cultured in vitro.A control group was established using HPAEC with blank serum,while a model group was created by stimulating HPAEC with 5 mg/L TGF-β1 for 72 hours.In the DAPT group,the same TGF-β1 stimulation was followed by the addition of 500 nmol/L Notch signalling inhibitor for 72 hours.Reverse transcription quantitative polymerase chain reaction(RT-PCR)was employed to measure the expression of Notch1,Jagged1,and the CBF1 Promoter(CBF1)mRNA,while Western blotting(WB)was used to assess the protein expressions of Notch1,Jagged1,and CBF1.Co-immunoprecipitation(Co-IP)was conducted to examine the binding of Notch1 intracellular domain protein(NICD1)with CBF1,and the Methylthiazoltetrazolium Colorimetry(MTT)assay was performed to evaluate cell proliferation.Transwell and scratch assays were conducted to measure cell migration ability,and immunofluorescence was used to detect the expression of endothelial cell markers CD31 and VE-cadherin,as well as mesenchymal cell markers fibroblast-specific protein 1(FSP1)and α-smooth muscle actin(α-SMA).Results Statistical analysis revealed significant differences(P<0.05)in the expression levels of Notch1,Jagged1,CBF1 mRNA and proteins,as well as cell proliferation and migration abilities among the three groups.The blank group and DAPT group exhibited lower expressions of Notch1,Jagged1,and CBF1 mRNA and proteins,alongside reduced cell proliferation and migration compared to the model group,with significant differences noted(P<0.05).Co-IP results indicated discrepancies in the binding of NICD1 and CBF1 among the three groups,with binding inhibited in the blank and DAPT groups,while binding occurred in the model group.Immunofluorescence results demonstrated differences in the expressions of CD31,VE-Cadherin,FSP1,and α-SMA among the 3 groups,where the blank and DAPT groups showed higher expressions of endothelial markers CD31 and VE-Cadherin compared to the model group,while FSP1 and α-SMA expressions were lower.Conclusion The Jagged1/Notch1 signalling pathway is involved in the EndMT process during IPF,and inhibiting this signalling pathway can suppress EndMT.关键词
特发性肺纤维化/内皮细胞/内皮间质转化/Jagged1/Notch1信号通路/人肺动脉内皮细胞Key words
Idiopathic pulmonary fibrosis/Endothelial cells/Endothelial-to-mesenchymal transition/Jagged1/Notch1 signaling pathway/Human pulmonary artery endothelial cells分类
医药卫生引用本文复制引用
杨其芬,赵惠亮,郭永胜,渠景连..Jagged1/Notch1信号通路对特发性肺纤维化过程中内皮间质转化的影响研究[J].中国全科医学,2025,28(25):3151-3160,10.基金项目
国家自然科学基金资助项目(81660789,82160892) （81660789,82160892）
