环境与职业医学2025，Vol.42Issue(2)：188-195,8.DOI:10.11836/JEOM24257
miR-130a-3p靶向PPAR-γ对肺上皮间质转化的影响在矽肺纤维化中的作用
Effect of miR-130a-3p targeting PPAR-γ on epithelial-mesenchymal transition in silica-induced pulmonary fibrosis
摘要
Abstract
[Background]At present,the treatment of silicosis is still limited,and no method is available to cure the disease.miRNAs are involved in the process of fibrosis at the transcriptional level by di-rectly degrading target gene mRNA or inhibiting its translation.However,how miR-130a-3p reg-ulates silicosis fibrosis has not been fully elucidated yet. [Objective]To investigate whether miR-130a-3p promotes epithelial-mesenchymal transition(EMT)by inhibiting peroxisome proliferators-activated receptors gamma(PPAR-γ),thereby pro-moting the process of silicotic fibrosis.To identify effective new targets for the treatment of silicotic fibrosis. [Methods](1)Animal experiments:C57BL/6J mice were intratracheally injected with a one-time dose of 10 mg silica suspension(dissolved in 100 μL saline)as positive lung exposure.A silicosis model group was established 28 d after the exposure.A control group was injected with the same amount of normal saline into the trachea.Hematoxylin-eosin staining and Sirius red staining were used to observe the pathological changes and collagen deposition in lung tissues respectively.Realtime fluorescence-based quantitative polymerase chain re-action(RT-qPCR)was used to assay the expression of miR-130a-3p and PPAR-γ mRNA in lung tissues.Western blotting was used to detect the protein expression of PPAR-γ,transforming growth factor(TGF)-β1,E-cadherin,α-smooth muscle actin(α-SMA),and Collagen Ⅰ in lung tissues.(2)Cells experiments:Mouse lung epithelial cells(MLE-12)were induced with 5 µg·L-1 TGF-β1 for different time(0,12,24,48 h).RT-qPCR was used to detect the expression of miR-130a-3p and PPAR-γ mRNA in cells.The binding relationship between miR-130a-3p and PPAR-γ mRNA was verified by dual luciferase reporter gene assay.MLE-12 cells were stimulated by 5 µg·L-1 TGF-β1 after transfection of miR-130a-3p inhibitor,and Western blotting was used to measure the protein expression of PPAR-γ,E-cadherin,and α-SMA in the TGF-β1-induced cells. [Results]In the silicosis model group,the alveolar septum was widened and the pulmonary nodules were formed.The Sirius red staining collagen deposition in pulmonary nodules indicated that a silicosis fibrosis model was successfully established.The expressions of TGF-β1,α-SMA,and Collagen Ⅰ proteins were increased,and the expressions of E-cadherin and PPAR-γ proteins were decreased in lung tissues of the silicosis group,compared with the control group(P＜0.05 or P＜0.01).The expression of miR-130a-3p was increased and the ex-pression of PPAR-γ mRNA was decreased in lung tissues of the silicosis model(P＜0.01).The expression of miR-130a-3p was significantly increased,while the expression of PPAR-γ mRNA was decreased in the TGF-β1 induced MLE-12 cells(P＜0.05 or P＜0.01).The dual luciferase reporter assay showed a direct relationship between miR-130a-3p and PPAR-γ mRNA in MLE-12 cells.The transfection of miR-130a-3p in-hibitor in the TGF-β1 induced MLE-12 cells inhibited the decrease of PPAR-γ and E-cadherin proteins,and the increase of α-SMA protein in the MLE-12 cells induced by TGF-β1(P＜0.05 or P＜0.01). [Conclusion]miR-130a-3p promotes the development of silicosis fibrosis by targeting PPAR-γ to increase pulmonary EMT.关键词
矽肺/miR-130a-3p/过氧化物酶增殖物激活受体γ/上皮间质转化/纤维化Key words
silicosis/miR-130a-3p/peroxisome proliferation activator receptor γ/epithelial-mesenchymal transition/fibrosis分类
预防医学引用本文复制引用
郝小惠,李倩,靳宜璇,张秦欣,王渝迪,杨方..miR-130a-3p靶向PPAR-γ对肺上皮间质转化的影响在矽肺纤维化中的作用[J].环境与职业医学,2025,42(2):188-195,8.基金项目
河北省自然科学基金项目(H202409025) （H202409025）
2021年河北省博士研究生创新资助项目(CXZZBS2021106) （CXZZBS2021106）
