P.g induces CKD by activating ferroptosis via the NF‑κB/NLRP3 pathway
After GMCs were cultured with MΦCM for 24 h (Fig. 6A and B), the CCK-8 and live/dead staining results showed that 12 μmol QNZ had no toxic effect on SV40-MES-13 cells and that QNZ protected GMCs from MΦCM (Fig. 7A and B). After the cells were pretreated with QNZ before being cocultured with MΦCM, the protein levels of NF-κB, NLRP3, ASC, Caspase 1 and IL-1β were downregulated (Fig. 7C and D, Supplementary Figure 5A, B). A similar change in the mRNA levels of NF-κB, NLRP3, ASC, Caspase 1 and IL-1β was observed (Fig. 7E and F). These results showed that activation of the NF‑κB/NLRP3 pathways in GMCs induced by the polarization of M1 macrophages was blocked by QNZ in vitro . More importantly, the protein levels of ACSL4, xCT and GPX4 indicated that ferroptosis activation in GMCs stimulated by MΦCM was alleviated by QNZ treatment (Fig. 7G, Supplementary Figure 5C). Similarly, the changes in the mRNA levels of GPX4, NCOA4 and PTGS2 in GMCs stimulated by MΦCM were reversed by QNZ (Fig. 7H). Moreover, QNZ alleviated the deposition of Fe2+ in the mitochondria of SV40-MES-13 cells, as shown by Fe2+ fluorescence staining (Fig. 6I). Therefore, these results indicated that P.g induced CKD by activating ferroptosis in GMCs after promoting the polarization of M1 macrophages, which depended on the NF‑κB/NLRP3 pathway.