Wogonin induces ferroptosis of rat CIA-FLS cells via NRF2/HO-1 signaling pathway.

AIM: To investigate the mechanism by which wogonin (WOG) induces ferroptosis in collagen-induced arthritis rat fibroblast-like synoviocytes (rat CIA-FLS cells) through the nuclear factor E2-related factor 2 (NRF2)/heme oxygenase-1 (HO-1) signaling pathway. METHODS: Rat CIA-FLS cells were divided into: control group, low, medium, and high dose of (25, 50 and 100 μmol/L) WOG group, ferroptosis inhibitor (LIP-1) group, LIP-1 + high dose WOG group, HO-1 agonist cobalt protoporphyrin (COPP) group, and COPP + high dose WOG group. CCK-8 assay was used for cell viability. Crystal violet staining was used for for cell morphology. The levels of oxidative stress markers glutathione (GSH), malondialdehyde (MDA), and superoxide dismutase (SOD) were measured. DCFH-DA fluorescent probe was used to detect the intracellular reactive oxygen species (ROS) content as well as Western blot to detect the protein expression levels of Kelch-like ECH-associated protein 1 (KEAP-1), NRF2 and HO-1. RESULTS: Compared with the normal control group, administration of WOG treatment resulted in a significant decrease in CIA-FLS cell viability (P< 0. 01, a significant increase in the level of oxidative stress (P<0. 01), a significant increase in the content of ROS (P< 0. 01), a significant decrease in the level of expression of NRF2 and HO-1 proteins (P<0. 01), and a significant increase in the level of KEAP-1 (P<0. 01) in the rat. Compared with the WOG group, the LIP-1-treated group showed a significant increase in cell viability (P<0. 01, a significant decrease in the level of oxidative stress (P<0. 01), and a significant decrease in the content of ROS (P<0. 01). Compared with the WOG group, the addition of COPP resulted in a significant increase in the protein expression levels of NRF2 and HO-1 (P<0. 01) and a significant decrease in KEAP-1 levels (P< 0.01). CONCLUSION: WOG can induce ferroptosis in rat CIA-FLS cells by promoting oxidative stress through the NRF2/HO-1 signaling pathway. [ABSTRACT FROM AUTHOR]