环境与职业医学2025，Vol.42Issue(12)：1429-1437,9.DOI:10.11836/JEOM25253
基于PGC-1α-NRF1-TFAM信号通路探究线粒体生物发生在煤工尘肺大鼠模型中的作用
Role of mitochondrial biogenesis in rat model of coal workers'pneumoconiosis based on PGC-1α-NRF1-TFAM signaling pathway
摘要
Abstract
[Background]Mitochondrial biogenesis is pivotal in coal workers'pneumoconiosis fibrosis,yet the role of the peroxisome proliferator-activated receptor gamma coactivator 1-alpha(PGC-1α)-nuclear respiratory factor 1(NRF1)-mitochondrial transcription factor A(TFAM)pathway in mitochondrial biogenesis remains elusive,warranting further investigation. [Objective]To elucidate the role of the PGC-1α-NRF1-TFAM pathway in mitochondrial biogenesis in a rat coal workers'pneumoconiosis model through in vivo and in vitro experiments. [Methods](1)n vivo:twelve SPF male SD rats(200-220 g)were randomized into a control group and a coal dust group(n=6 per group).After acclimatization,the coal dust group received 1 mL 50 mg·mL-1 coal dust suspension via intratracheal instillation;the controls received saline.Lung tissues were harvested after two months for histopathology[HE,Masson,and transmission electron microscopy(TEM)],protein and mRNA analysis,and mitochondrial DNA(mtDNA)quantification by quantitative real-time polymerase chain reaction(qPCR).(2)In vitro:rat lung type II epithelial cells(RLE-6TN)cells were exposed to coal dust(50,100,200,and 400 mg·L-1,24 h).CCK-8 assay de-termined optimal doses.Ultrastructural changes were analyzed by TEM.Cells were transfected with OE-PGC-1α(PGC-1α overexpression)or shRNA-PGC-1α plasmids(PGC-1α knockdown),and the transfection efficiency was determined by reverse transcription quantitative real-time polymerase chain reaction(RT-qPCR).The expression levels of alpah-smooth muscle actin(α-SMA),citrate synthase(CS),PGC-1α,NRF1,TFAM,and fibronectin(Fn)proteins and their corresponding mRNA were detected using Western blot and RT-qPCR,respec-tively.The relative content of mtDNA was determined by qPCR. [Results]In vivo:the control group lung samples exhibited soft,pink parenchyma,while the coal dust-exposed lungs showed blackened surfaces with soft texture.The histopathological evaluation revealed intact alveolar walls in the controls versus structural destruction,micro-nodules,and fibrotic areas in the coal dust group.After Masson staining,coal dust deposits were found surrounded by blue collagen fibers in the exposed lungs,but absent in the controls.The coal dust group displayed significant upregulation of fibrotic marker α-SMA and downregulation of mitochondrial biogenesis markers(CS,PGC-1α,NRF1,TFAM)and mtDNA compared to the controls(P<0.05).In vitro:coal dust exposure reduced cell density and induced morphological alterations.TEM revealed evenly distributed normal mitochondria in controls versus mitochondrial swelling,disrupted cristae,and reduced numbers in exposed cells.The mitochondrial biogenesis markers were elevated in the coal dust+OE-PGC-1α group compared to the coal dust+OE-NC group(P<0.05);in contrast,they were decreased in the coal dust+shRNA-PGC-1α group compared to the coal dust+shRNA-NC group(P<0.05).Compared to the control group,the ex-pression levels of the fibrosis marker α-SMA mRNA and protein were increased in the coal dust group(P<0.05).Overexpression of PGC-1α reduced α-SMA expression,while downregulation of PGC-1α increased its expression(P<0.05). [Conclusion]Coal dust exposure induces mitochondrial dysfunction and pulmonary fibrosis in vivo and in vitro via the PGC-1α-NRF1-TFAM pathway dysregulation.Targeting this pathway may mitigate coal dust-induced fibrosis by restoring mitochondrial biogenesis.关键词
煤尘/纤维化/线粒体生物发生/过氧化物酶体增殖物激活受体-γ共激活因子-1αKey words
coal dust/fibrosis/mitochondrial biogenesis/peroxisome proliferator-activated receptor gamma coactivator 1-alpha分类
医药卫生引用本文复制引用
章梅,韩晓强,刘琭璐,王燕,马鑫,熊育,杨惠芳,张娜..基于PGC-1α-NRF1-TFAM信号通路探究线粒体生物发生在煤工尘肺大鼠模型中的作用[J].环境与职业医学,2025,42(12):1429-1437,9.基金项目
国家自然科学基金项目(82260636,82160608) （82260636,82160608）
宁夏自然科学基金项目(2021BEB04059,2021AAC02013,2022AAC03141) （2021BEB04059,2021AAC02013,2022AAC03141）
