线粒体分裂与氧化应激的相互促进作用导致了百草枯诱导的肺纤维化中线粒体DNA介导的炎症和上皮-间质转化。
Mutual promotion of mitochondrial fission and oxidative stress contributes to mitochondrial-DNA-mediated inflammation and epithelial-mesenchymal transition in paraquat-induced pulmonary fibrosis.
作者信息
Zhang Jie, Li Wen-Jing, Chen Shi-Qiang, Chen Ze, Zhang Chen, Ying Ran, Liu Hong-Bing, Chen Long-Wang, Tang Ya-Hui, Lu Zhong-Qiu, Zhao Guang-Ju
机构信息
Department of Emergency Medicine, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China.
Wenzhou Key Laboratory of Emergency and Disaster Medicine, Wenzhou 325000, China.
出版信息
World J Emerg Med. 2023;14(3):209-216. doi: 10.5847/wjem.j.1920-8642.2023.057.
BACKGROUND
Pulmonary fibrosis (PF) is one of the main causes of death in patients with paraquat (PQ) poisoning. This study aimed to evaluate the relationship between mitochondrial fission and oxidative stress in PQ-induced epithelial-mesenchymal transition (EMT) and PF.
METHODS
C57BL/6 mice and MLE-12 cells were exposed to PQ to construct a PF model and . Histological changes in the lungs were examined by hematoxylin and eosin (H&E) staining. Mitochondrial morphology was detected by MitoTracker Deep Red FM or transmission electron microscopy (TEM). Western blotting and immunofluorescence were used to determine the expression of protein. The migration ability of the cells was detected by the cell scratch test. Mitochondrial DNA (mtDNA) levels were assessed by real-time polymerase chain reaction (PCR). Enzyme-linked immunosorbent assay (ELISA) was applied to detect cytokine levels. Superoxide dismutase (SOD) activity and the levels of glutathione (GSH) and malondialdehyde (MDA) were detected by chemichromatometry.
RESULTS
PQ exposure caused EMT and PF and . PQ destroyed mitochondrial structure and enhanced the expression of dynamin-related protein 1 (Drp1), which were accompanied by oxidative stress. Inhibiting mitochondrial fission using mitochondrial division inhibitor-1 (Mdivi-1), a selective inhibitor of Drp1, attenuated PQ-induced EMT and oxidative damage. Treatment with N-acetyl-L-cysteine (NAC), an antioxidant, reduced Drp1 expression, attenuated mitochondrial structure damage and inhibited PQ-induced EMT and PF. Both Mdivi-1 and NAC treatment markedly suppressed mtDNA release, the expression of Toll-like receptor 9 (TLR9) and phosphorylation (P)-NF-κB p65 as well as cytokines (interleukin 6 [IL-6], interleukin-1β [IL-1β], and tumor necrosis factor-α [TNF-α]) production.
CONCLUSION
Mutual promotion of mitochondrial fission and oxidative stress contributes to EMT in PQ-induced PF, which is associated with the mtDNA/TLR9/NF-κB pathway.
背景
肺纤维化（PF）是百草枯（PQ）中毒患者的主要死因之一。本研究旨在评估PQ诱导的上皮-间质转化（EMT）和PF中线粒体分裂与氧化应激之间的关系。
方法
将C57BL/6小鼠和MLE-12细胞暴露于PQ以构建PF模型。通过苏木精和伊红（H&E）染色检查肺组织学变化。用MitoTracker Deep Red FM或透射电子显微镜（TEM）检测线粒体形态。采用蛋白质印迹法和免疫荧光法测定蛋白质表达。通过细胞划痕试验检测细胞的迁移能力。通过实时聚合酶链反应（PCR）评估线粒体DNA（mtDNA）水平。应用酶联免疫吸附测定（ELISA）检测细胞因子水平。通过化学比色法检测超氧化物歧化酶（SOD）活性以及谷胱甘肽（GSH）和丙二醛（MDA）水平。
结果
PQ暴露导致EMT和PF。PQ破坏线粒体结构并增强动力相关蛋白1（Drp1）的表达，同时伴有氧化应激。使用线粒体分裂抑制剂-1（Mdivi-1，一种Drp1的选择性抑制剂）抑制线粒体分裂，可减轻PQ诱导的EMT和氧化损伤。用抗氧化剂N-乙酰半胱氨酸（NAC）处理可降低Drp1表达，减轻线粒体结构损伤，并抑制PQ诱导的EMT和PF。Mdivi-1和NAC处理均显著抑制mtDNA释放、Toll样受体9（TLR9）的表达、磷酸化（P）-NF-κB p65以及细胞因子（白细胞介素6 [IL-6]、白细胞介素-1β [IL-1β]和肿瘤坏死因子-α [TNF-α]）的产生。
结论
线粒体分裂与氧化应激相互促进，导致PQ诱导的PF中的EMT，这与mtDNA/TLR9/NF-κB途径有关。
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