Ferrostatin‑1 alleviates oxalate‑induced renal tubular epithelial cell injury, fibrosis and calcium oxalate stone formation by inhibiting ferroptosis

Xie,Jinna; Ye,Zehua; Li,Lei; Xia,Yuqi; Yuan,Run; Ruan,Yuan; Zhou,Xiangjun

doi:10.3892/mmr.2022.12772

Ferrostatin‑1 alleviates oxalate‑induced renal tubular epithelial cell injury, fibrosis and calcium oxalate stone formation by inhibiting ferroptosis

Authors:
- Jinna Xie
- Zehua Ye
- Lei Li
- Yuqi Xia
- Run Yuan
- Yuan Ruan
- Xiangjun Zhou
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Affiliations: Department of Urology, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China
Published online on: June 15, 2022 https://doi.org/10.3892/mmr.2022.12772
Article Number: 256
Copyright: © Xie et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The present study aimed to evaluate the role and mechanism of ferrostatin‑1 (Fer‑1) in oxalate (Ox)‑induced renal tubular epithelial cell injury, fibrosis, and calcium oxalate (CaOx) stone formation. A CaOx model in mice kidneys was established via intraperitoneal injection of 80 mg/kg glyoxylic acid for 14 days. The mice were randomly divided into three groups (n=6), namely, the control (Con), the CaOx group, and the CaOx + Fer‑1 group. Cultured human renal tubular epithelial cells (HK‑2 cells) were randomly divided into three groups (n=3), namely, the control (Con), the Ox group, and the Ox + Fer‑1 group. The levels of heme oxygenase 1 (HO‑1), superoxide dismutase 2 (SOD2), glutathione peroxidase 4 (GPX4), and solute carrier family 7 member 11 (SLC7A11) were assessed by immunofluorescence and western blot analysis. Renal tubular injury and apoptosis were evaluated by H&E and TUNEL staining. Kidney interstitial fibrosis was evaluated by Masson and Sirius red staining, and the levels of E‑cadherin, vimentin and α‑SMA were detected by immunofluorescence or western blot analysis. Mitochondrial structure was observed using a transmission electron microscope. The levels of reactive oxygen species (ROS) were determined by flow cytometry and CaOx stone formation was evaluated by von Kossa staining. The results revealed that in comparison with the Con group, mitochondrial injury under glyoxylic acid treatment was observed by TEM. The expression of GPX4 and SLC7A11 in the CaOx and Ox groups was downregulated (P<0.05), whereas the expression of HO‑1 and SOD2 was upregulated (P<0.05). Renal tissue damage, apoptosis of renal tubular epithelial cells, and interstitial fibrosis were increased in the CaOx and Ox groups (P<0.05). In comparison with the CaOx or Ox group, the expression of GPX4 and SLC7A11 in the CaOx + Fer‑1 or Ox + Fer‑1 group was upregulated (P<0.05), whereas that of HO‑1 and SOD2 was downregulated (P<0.05). Renal tissue damage, apoptosis of renal tubular epithelial cells and interstitial fibrosis were decreased following Fer‑1 treatment (P<0.05). The ROS level was also decreased following Fer‑1 treatment. Moreover, CaOx stone formation was decreased in the CaOx + Fer‑1 group (P<0.05). In conclusion, Fer‑1 alleviated Ox‑induced renal tubular epithelial cell injury, fibrosis, and CaOx stone formation by inhibiting ferroptosis.

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