Iso-seco-tanapartholide from Artemisia argyi inhibits the PFKFB3-mediated glycolytic pathway to attenuate airway inflammation in lipopolysaccharide-induced acute lung injury mice

In traditional Chinese folk medicine, Artemisia argyi H.Lév. & Vaniot (A. argyi) has been used for thousands of years, and it is clinically used to treat bronchitis and asthma. However, the mechanism of action of A. argyi on respiratory tract inflammation is not clear. Accumulating evidence that phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) is actively expressed in inflammation. Here, we found that iso-seco-tanapartholide (IST), a sesquiterpene isolated from A. argyi, exhibited potent anti-inflammatory activity and significant inhibition of PFKFB3 expression. Therefore, we evaluated the effect of IST on airway inflammation and revealed its possible mechanisms.This study aimed to investigate the protective effect and possible mechanism of IST in lipopolysaccharide (LPS)-induced acute lung injury in mice.In vitro, RAW264.7 cells and BMDMs were stimulated with LPS, and the level of NO and inflammatory factors TNF-α, IL-1β, and IL-6 were detected by Griess reagent and ELISA, respectively. The effect of IST on the levels of PFKFB3 and its downstream proteins (p-STAT3, p-p65) in cells was assayed by western blotting. Lactate and glycolytic phenotypes were detected by lactate kit and Seahorse assay. In vivo, a mouse model of acute lung injury was induced by LPS, and the levels of inflammatory factors were measured by ELISA. Expression of PFKFB3 and its downstream proteins (p-STAT3, p-p65) in mouse alveolar macrophages by western blotting analysis. Lung permeability assessment by Evans Blue dye assay. H&E staining and Immunocytochemistry were used to observe the protection of IST against lung injury.IST significantly reduced LPS-induced expression of PFKFB3 and its downstream proteins (p-STAT3, p-p65). The inhibition of PFKFB3 has an impact on the glycolytic phenotype, such as a reduction in the rate of extracellular acidification (ECAR) and elevated lactate levels, and an increase in the rate of cellular oxygen consumption (OCR). Furthermore, IST inhibited LPS-induced NO release and increased the expression of pro-inflammatory factors TNF-α, IL-1β, and IL-6. In vivo, IST reduced pulmonary edema in LPS-induced acute lung injury, improved lung function, and reduced levels of inflammatory factors and lactate secretion.These results suggest that IST improves the characteristics of ALI by inhibiting the expression of the PFKFB3-mediated glycolytic pathway and may be a potential anti-inflammatory agent for inflammation-related lung diseases.