Fig. 8

ARND manifested an anti-fibrotic effect. A Calu-3 cells were treated with different concentrations of ARND (0.625–5 µL/mL) for 48 h. Treatment with different concentrations of ARND did not reduce viability in Calu-3 cells, indicating ARND manifested no substantial toxicity towards Calu-3 cells. (n = 4). B Calu-3 cells were exposed to different concentrations of H₂O₂ for 24 h. A decline in cell viability was observed at H₂O₂ concentrations between 62.5 (p < 0.05) and 125 µM (p < 0.001). Therefore, the approximate mean of the two concentrations i.e., 100 µM H₂O₂ was selected for establishing the cell model of lung fibrosis (n = 3). *p < 0.05 and ***p < 0.001 vs 0 µM. C–D The mRNA expression levels of 10 biomarkers related to epithelial-mesenchymal transition (EMT; TGF-β, CXCL-16, CXCR4, IL-17, IL-25 and α-SMA) as well as extracellular matrix (ECM; Col-1a, Col-4a, elastin, and Fn1) of fibrosis were examined after ARND pre-treatment and co-treatment with H₂O₂. The group without receiving any treatment served as the control group while the group with only H₂O₂ (100 µM) treatment served as the model group. There was a significant upregulation of mRNA expression levels of the 10 fibrotic biomarkers in the model group (p < 0.05), while cells co-treated with ARND demonstrated a significant downregulation of the mRNA expressions levels of different fibrotic biomarkers, albeit to different extents (p < 0.05). (n = 4). **p < 0.01 and ***p < 0.001 vs the control group; ^#p < 0.05, ^##p < 0.01 and ^###p < 0.001 vs the model group