Ophiopogonin D attenuates PM2.5-induced inflammation via suppressing the AMPK/NF-κB pathway in mouse pulmonary epithelial cells

Exposure to fine particulate matter, such as particulate matter of ≤2.5 µm in diameter (PM2.5), causes pulmonary inflammation and injury to other organs. It has been reported that Ophiopogonin D (OP-D) has anti-inflammatory activity. The aim of the present study was to investigate this anti-inflammatory activity of OP-D on PM2.5-induced acute airway inflammation and its underlying mechanisms. The viability of PM2.5-treated mouse lung epithelial (MLE-12) cells with or without OP-D treatment was determined using a Cell Counting Kit-8 assay. The corresponding levels of IL-1β, IL-6, IL-8 and TNF-α were examined via ELISA. Subcellular localization of NF-κBp65 was detected using immunofluorescence staining. The expression levels of AMP-activated protein kinase (AMPK), phosphorylated (p)-AMPK, NF-κBp65 and p-NF-κBp65 were analyzed using western blotting. The selective AMPK inhibitor Compound C (CC) was utilized to investigate the involvement of AMPK in the protection against PM2.5-induced cell inflammation by OP-D treatment. The results demonstrated that OP-D significantly ameliorated the PM2.5-stimulated release of proinflammatory cytokines (TNF-α, IL-1β, IL-6 and IL-8) and inhibited the translocation of NF-κBp65 from the cytoplasm to the nucleus in MLE-12 cells. Moreover, OP-D significantly prevented the PM2.5-triggered phosphorylation of NF-κBp65 and upregulated AMPK activity. The anti-inflammatory activity of OP-D could also be attenuated by the AMPK-specific inhibitor CC. The present results suggested that the anti-inflammatory activity of OP-D was mediated via AMPK activation and NF-κB signaling pathway downregulation, which ameliorated the expression of proinflammatory cytokines. Therefore, OP-D could be a candidate drug to treat PM2.5-induced airway inflammation.