Andrographolide attenuates epithelial-mesenchymal transition induced by TGF-β1 in alveolar epithelial cells

Abstract

Andrographolide (Andro), a component from Chinese medicinal herb Andrographis paniculata, could alleviate pulmonary fibrosis in rodents. Yet, whether and how Andro mitigates epithelial-mesenchymal transition (EMT) induced by TGF-β1 remain unknown. This study aimed to explore the effect of Andro on TGF-β1-induced EMT in human alveolar epithelial cells (AECs) and the mechanisms involved. We illustrated that Andro inhibited TGF-β1-induced EMT and EMT-related transcription factors in alveolar epithelial A549 cells. Andro also reduced TGF-β1-induced cell migration and synthesis of pro-fibrotic factors (ie CCN-2, TGF-β1), matrix metalloproteinases (ie MMP-2, MMP-9) and extracellular matrix (ECM) components (ie collagen 1), implying the inhibiting effect of Andro on TGF-β1-induced EMT-like cell behaviours. Mechanistically, Andro treatment not only repressed TGF-β1-induced Smad2/3 phosphorylation and Smad4 nuclear translocation, but also suppressed TGF-β1-induced Erk1/2 phosphorylation and nuclear translocation in A549 cells. And treatment with ALK5 inhibitor (SB431542) or Erk1/2 inhibitors (SCH772984 and PD98059) remarkably reduced EMT evoked by TGF-β1. In addition, Andro also reduced TGF-β1-induced intracellular ROS generation and NOX4 expression, and elevated antioxidant superoxide dismutase 2 (SOD2) expression, demonstrating the inhibiting effect of Andro on TGF-β1-induced oxidative stress, which is closely linked to EMT. Furthermore, Andro remarkably attenuated TGF-β1-induced down-regulation of sirtuin1 (Sirt1) and forkhead box O3 (FOXO3), implying that Andro protects AECs from EMT partially by activating Sirt1/FOXO3-mediated anti-oxidative stress pathway. In conclusion, Andro represses TGF-β1-induced EMT in AECs by suppressing Smad2/3 and Erk1/2 signalling pathways and is also closely linked to the activation of sirt1/FOXO3-mediated anti-oxidative stress pathway.

Keywords: EMT; Sirt1/FOXO3 signalling; TGF-β1; alveolar epithelial cells; andrographolide; oxidative stress.