Figure 1

Plumbagin inhibits aerobic glycolysis in activated macrophages. (A) Bone marrow-derived macrophages (BMDMs) were treated with plumbagin (1 and 3 µmol/L) for 24 h, and cell viability was analyzed. (B–C) BMDMs and peritoneal macrophages (PMs) were pretreated with plumbagin and 2DG for 1 h and then stimulated with lipopolysaccharide (LPS) (100 ng/mL) for 2 or 24 h. After the drug treatment ended, the cells were changed to fresh culture medium and then the oxygen consumption rates (OCR, indicative of oxidative phosphorylation) and extracellular acidification rates (ECAR, indicative of glycolysis) were monitored using the Mito Stress Test Kit (Oligomycin “Olg,” 100 µmol/L; FCCP, 100 µmol/L; Rotenone/antimycin A “A/R,” 50 µmol/L) and Glycolysis Stress Test Kit (Glucose “Glu,” 10 mM; Oligomycin “Olg,” 1 µmol/L; 2-DG, 50 µmol/L) by Seahorse Bioscience Extracellular Flux Analyzer. (D) In parallel, phosphoenolpyruvate (PEP) and lactate levels were assayed using commercial kits (n = 3, *, p < 0.05 versus LPS group).