Qiweizhigan granule ameliorates metabolic dysfunction-associated steatohepatitis by modulating the galectin 3/tumor necrosis factor receptor-associated factor 6-mediated ferroptosis

Figure 7 Qiweizhigan granule ameliorates metabolic dysfunction-associated steatohepatitis by modulating the galectin 3/tumor necrosis factor receptor-associated factor 6/ glutathione peroxidase 4 axis. A: The levels of tumor necrosis factor receptor-associated factor 6 (TRAF6) and NOD-like receptor family pyrin domain containing 3 (NLRP3) in RAW264.7 cells after galectin 3 (LGALS3) knockdown were performed; B: The levels of TRAF6 and NLRP3 in RAW264.7 cells after LGALS3 overexpression were performed; C and D: The levels of glutathione peroxidase 4, NLRP3, and TRAF6 in RAW264.7 cells after LGALS3 overexpression with subsequent TRAF6 inhibition by C25-140 were verified; E: Measurement of total iron content in RAW264.7 cells after LGALS3 overexpression combined with TRAF6 inhibitor treatment; F: Measurement of ferrous iron (Fe²⁺) content in RAW264.7 cells after LGALS3 overexpression combined with TRAF6 inhibitor treatment; G and H: The mRNA expression of TRAF6 and NLRP3 in liver tissues among Normal, choline-deficient, L-amino acid-defined high-fat diet (CDAHFD), and Qiweizhigan granule (QWZG) groups were performed; I: Immunohistochemical analysis of TRAF6 in liver sections among the Normal, CDAHFD, and QWZG groups was performed; J: Immunofluorescence analysis of NLRP3 in liver sections among the Normal, CDAHFD, and QWZG groups was performed; K: Protein levels of TRAF6 and NLRP3 in liver tissues among the Normal, CDAHFD, and QWZG groups were performed. Data were presented as means ± SEM. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. CDAHFD: Choline-deficient, L-amino acid-defined high-fat diet; GPX4: Glutathione peroxidase 4; LGALS3: Galectin 3; NLRP3: NOD-like receptor family pyrin domain containing 3; QWZG: Qiweizhigan granule; TRAF6: Tumor necrosis factor receptor-associated factor 6.