Department of Applied Biology, CSIR-Indian Institute of Chemical Technology (IICT), Hyderabad 500 007, India.
Department of Applied Biology, CSIR-Indian Institute of Chemical Technology (IICT), Hyderabad 500 007, India.
Phytomedicine. 2020 Nov;78:153298. doi: 10.1016/j.phymed.2020.153298. Epub 2020 Aug 1.
Idiopathic Pulmonary Fibrosis (IPF) is a progressive inflammatory disorder driven by a fibrotic cascade of events such as epithelial to mesenchymal transition, extracellular matrix production and collagen formation in the lungs in a sequential manner. IPF incidences were raising rapidly across the world. FDA approved pirfenidone and nintedanib (tyrosine kinase inhibitors) are being used as a first-line treatment drugs for IPF, however, neither the quality of life nor survival rates have been improved because of patient noncompliance due to multiple side effects. Thus, the development of novel therapeutic approaches targeting TGF-β mediated cascade of fibrotic events is urgently needed to improve the survival of the patients suffering from devastating disease.
The aim of this study was to investigate and validate the anti-fibrotic properties of Biochanin-A (isoflavone) against TGF-β mediated fibrosis in in vitro, ex vivo, in vivo models and to determine the molecular mechanisms that mediate these anti-fibrotic effects.
The therapeutic activity of BCA was determined in in vitro/ex vivo models. Cells were pre-treated with BCA and incubated in presence or absence of recombinant-TGF-β to stimulate the fibrotic cascade of events. Pulmonary fibrosis was developed by intratracheal administration of bleomycin in rats. BCA treatment was given for 14 days from post bleomycin instillation and then various investigations (collagen content, fibrosis gene/protein expression and histopathological changes) were performed to assess the anti-fibrotic activity of BCA.
In vitro/ex vivo (Primary normal, IPF cell line and primary IPF cells/ Precision cut mouse lung slices) experiments revealed that, BCA treatment significantly (p < 0.001) reduced the expression of TGF-β modulated fibrotic genes/protein expressions (including their functions) which are involved in the cascade of fibrotic events. BCA treatment significantly (p < 0.01) reduced the bleomycin-induced inflammatory cell-infiltration, inflammatory markers expression, collagen deposition and expression of fibrotic markers in lung tissues equivalent or better than pirfenidone treatment. In addition, BCA treatment significantly (p < 0.001) attenuated the TGF-β1/BLM-mediated increase of TGF-β/Smad2/3 phosphorylation and resulted in the reduction of pathological abnormalities in lung tissues determined by histopathology observations.
Collectively, BCA treatment demonstrated the remarkable therapeutic effects on TGF-β/BLM mediated pulmonary fibrosis using IPF cells and rodent models. This current study may offer a novel treatment approach to halt and may be even rescue the devastating lung scarring of IPF.
特发性肺纤维化（IPF）是一种进行性炎症性疾病，其特征是上皮细胞向间充质转化、细胞外基质产生和胶原形成等纤维化级联反应依次发生。特发性肺纤维化的发病率在全球范围内迅速上升。美国食品和药物管理局（FDA）批准的吡非尼酮和尼达尼布（酪氨酸激酶抑制剂）被用作 IPF 的一线治疗药物，然而，由于多种副作用导致患者不遵医嘱，生活质量和生存率都没有得到改善。因此，迫切需要开发针对 TGF-β 介导的纤维化级联反应的新型治疗方法，以提高患有这种破坏性疾病的患者的生存率。
本研究旨在探讨和验证生物黄酮 A（异黄酮）在体外、离体、体内模型中对 TGF-β 介导的纤维化的抗纤维化作用，并确定介导这些抗纤维化作用的分子机制。
在体外/离体模型中测定 BCA 的治疗活性。细胞先用 BCA 预处理，然后在存在或不存在重组-TGF-β的情况下孵育，以刺激纤维化级联反应。通过气管内给予博来霉素在大鼠中诱导肺纤维化。博来霉素给药后 14 天给予 BCA 治疗，然后进行各种研究（胶原含量、纤维化基因/蛋白表达和组织病理学变化），以评估 BCA 的抗纤维化活性。
体外/离体（原代正常、IPF 细胞系和原代 IPF 细胞/精密切割鼠肺切片）实验表明，BCA 治疗显著（p < 0.001）降低了 TGF-β 调节的纤维化基因/蛋白表达（包括其功能），这些基因/蛋白表达参与了纤维化级联反应。BCA 治疗显著（p < 0.01）减少了博来霉素诱导的炎症细胞浸润、炎症标志物表达、胶原沉积和肺组织中纤维化标志物的表达，其效果与吡非尼酮治疗相当或更好。此外，BCA 治疗显著（p < 0.001）减弱了 TGF-β1/BLM 介导的 TGF-β/Smad2/3 磷酸化的增加，并导致通过组织病理学观察确定的肺组织中病理异常的减少。
综上所述，BCA 治疗在使用 IPF 细胞和啮齿动物模型的 TGF-β/BLM 介导的肺纤维化中表现出显著的治疗效果。本研究可能为阻止甚至可能挽救 IPF 的破坏性肺纤维化提供一种新的治疗方法。
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