Fig. 1

Homocysteine impaired β-catenin signaling and promoted ferroptosis in renal tubular epithelial cells. The hyperhomocysteinemia model was established by administering drinking water containing 1.8 g/L homocysteine to the mice. In the in vitro experiments, HKC-8 cells were treated with homocysteine at a concentration of 500 µmol/L. (A) Western blotting was employed to evaluate the expression levels of active β-catenin, β-catenin, GPX4, and FTH1 in the kidneys of mice. (B–E) Quantitative analysis was conducted to determine the levels of active β-catenin, β-catenin, GPX4, and FTH1 in the kidneys of mice. (F–H) Biochemical assays were performed to measure the levels of Fe²⁺, MDA, and GSH in mouse kidney tissues. (I) The morphology of mitochondria in renal tubular epithelial cells was examined using transmission electron microscopy. Yellow arrows highlighted mitochondrial damage, including the rupture of the mitochondrial membrane and the reduction of mitochondrial cristae. Scale bar, 300 nm. (J) Western blotting was employed to evaluate the expression levels of active β-catenin, β-catenin, GPX4, and FTH1 in HKC-8. (K–N) Quantitative analysis was conducted to determine the levels of active β-catenin, β-catenin, GPX4, and FTH1 in HKC-8. (O–Q) Biochemical assays were performed to measure the levels of Fe²⁺, MDA, and GSH in HKC-8. (R) Mitochondrial morphology in HKC-8 cells was examined using electron microscopy. Yellow arrows highlighted mitochondrial damage, including the rupture of the mitochondrial membrane and the reduction of mitochondrial cristae. Scale bar, 300 nm. *P < 0.05 vs. the controls (n = 6). Hcy, homocysteine; TEM, transmission electron microscopy.