Yenepoya Research Centre, Yenepoya University (Deemed to be University), Deralakatte, Mangalore 575018, Karnataka, India.
Comprehensive Pneumology Center, Helmholtz-Zentrum, Max-Lebsche-Platz-31, München 81377, Germany.
J Proteome Res. 2020 Aug 7;19(8):2950-2963. doi: 10.1021/acs.jproteome.9b00838. Epub 2020 Jul 22.
Bleomycin (BLM)-induced pulmonary fibrosis is characterized by inflammation in the alveoli, subsequent deposition of extracellular matrix (ECM) and myofibroblasts, and an impaired fibrinolytic system. Here, we describe major hematological changes, the IL-17A-mediated p53-fibrinolytic pathway, and the high throughput hits of liquid chromatography-mass spectrometry (LC-MS) analysis during the progression of pulmonary fibrosis and the therapeutic potential of curcumin against disease progression. C57BL/6 mice were exposed to BLM, followed by curcumin intervention after 24 and 48 h. Mice were sacrificed after 7 days to validate the hematological parameters, molecular pathways, and proteomics. Various techniques such as western blotting, immunofluorescence, reverse transcriptase polymerase chain reaction (RT-PCR), hematoxylin and eosin staining, Masson's trichrome staining, and immunohistochemistry were used to validate the proposed theory. LC-MS analysis was performed using a Q-Orbitrap mass spectrometer. The Schrödinger approach was used to perform the molecular docking studies. BLM-exposed mice exhibited gradual weight loss and altered lung morphology; however, these were reversed by curcumin treatment. Significant changes in the hematological parameters confirmed the severity of BLM exposure in the mice, and expression of IL-17A-mediated p53-fibrinolytic system components and alveolar epithelial cell (AEC) apoptosis further confirmed the pathophysiology of pulmonary fibrosis. Differentially expressed proteins were characterized and mapped using the proteomics approach. A strong interaction of curcumin is observed with p53, uPA, and PAI-I proteins. The key role of IL-17A-mediated inflammation in the impairment of the p53-fibrinolytic system and AEC apoptosis was confirmed during BLM-induced pulmonary fibrosis. Therapeutic efficacy of curcumin exhibited a protective role against the progression of pulmonary fibrosis, which promises potent therapeutic modality to target the IL-17A-mediated p53-fibrinolytic system during pulmonary fibrosis.
博莱霉素（BLM）诱导的肺纤维化的特征是肺泡炎症、细胞外基质（ECM）和肌成纤维细胞的沉积以及纤溶系统受损。在这里，我们描述了主要的血液学变化、IL-17A 介导的 p53 纤溶途径以及在肺纤维化进展过程中的液相色谱-质谱分析（LC-MS）高通量命中以及姜黄素对疾病进展的治疗潜力。C57BL/6 小鼠暴露于 BLM 后，在 24 和 48 小时后用姜黄素干预。在 7 天后处死小鼠以验证血液学参数、分子途径和蛋白质组学。使用 Western blot、免疫荧光、逆转录聚合酶链反应（RT-PCR）、苏木精和伊红染色、Masson 三色染色和免疫组织化学等各种技术来验证所提出的理论。使用 Q-Orbitrap 质谱仪进行 LC-MS 分析。Schrödinger 方法用于进行分子对接研究。暴露于 BLM 的小鼠表现出逐渐的体重减轻和改变的肺形态；然而，这些变化在姜黄素治疗后得到了逆转。血液学参数的显著变化证实了 BLM 暴露在小鼠中的严重程度，并且 IL-17A 介导的 p53 纤溶系统成分和肺泡上皮细胞（AEC）凋亡的表达进一步证实了肺纤维化的病理生理学。使用蛋白质组学方法对差异表达的蛋白质进行了表征和映射。姜黄素与 p53、uPA 和 PAI-I 蛋白之间存在强烈的相互作用。在 BLM 诱导的肺纤维化中，IL-17A 介导的炎症在 p53 纤溶系统和 AEC 凋亡的损伤中起关键作用。姜黄素的治疗功效表现出对肺纤维化进展的保护作用，这有望成为针对肺纤维化期间 IL-17A 介导的 p53 纤溶系统的有效治疗方式。
