Biomolecules & Therapeutics

The Korean Society of Applied Pharmacology (한국응용약물학회)
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Synergistic Renoprotective Effect of Melatonin and Zileuton by Inhibition of Ferroptosis via the AKT/mTOR/NRF2 Signaling in Kidney Injury and Fibrosis

  • Kyung Hee Jung (Department of Medicine, College of Medicine, and Program in Biomedical Science & Engineering, Inha University) ;
  • Sang Eun Kim (Department of Medicine, College of Medicine, and Program in Biomedical Science & Engineering, Inha University) ;
  • Han Gyeol Go (Department of Medicine, College of Medicine, and Program in Biomedical Science & Engineering, Inha University) ;
  • Yun Ji Lee (Department of Medicine, College of Medicine, and Program in Biomedical Science & Engineering, Inha University) ;
  • Min Seok Park (Department of Medicine, College of Medicine, and Program in Biomedical Science & Engineering, Inha University) ;
  • Soyeon Ko (Department of Medicine, College of Medicine, and Program in Biomedical Science & Engineering, Inha University) ;
  • Beom Seok Han (Department of Medicine, College of Medicine, and Program in Biomedical Science & Engineering, Inha University) ;
  • Young-Chan Yoon (Department of Medicine, College of Medicine, and Program in Biomedical Science & Engineering, Inha University) ;
  • Ye Jin Cho (Department of Medicine, College of Medicine, and Program in Biomedical Science & Engineering, Inha University) ;
  • Pureunchowon Lee (Department of Medicine, College of Medicine, and Program in Biomedical Science & Engineering, Inha University) ;
  • Sang-Ho Lee (Division of Nephrology, Department of Internal Medicine, College of Medicine, Kyung Hee University) ;
  • Kipyo Kim (Divison of Nephrology and Hypertension, Department of Internal Medicine, Inha University Hospital, Inha University College of Medicine) ;
  • Soon-Sun Hong (Department of Medicine, College of Medicine, and Program in Biomedical Science & Engineering, Inha University)
  • Received : 2023.03.23
  • Accepted : 2023.04.17
  • Published : 2023.11.01
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Abstract

According to recent evidence, ferroptosis is a major cell death mechanism in the pathogenesis of kidney injury and fibrosis. Despite the renoprotective effects of classical ferroptosis inhibitors, therapeutic approaches targeting kidney ferroptosis remain limited. In this study, we assessed the renoprotective effects of melatonin and zileuton as a novel therapeutic strategy against ferroptosis-mediated kidney injury and fibrosis. First, we identified RSL3-induced ferroptosis in renal tubular epithelial HK-2 and HKC-8 cells. Lipid peroxidation and cell death induced by RSL3 were synergistically mitigated by the combination of melatonin and zileuton. Combination treatment significantly downregulated the expression of ferroptosis-associated proteins, 4-HNE and HO-1, and upregulated the expression of GPX4. The expression levels of p-AKT and p-mTOR also increased, in addition to that of NRF2 in renal tubular epithelial cells. When melatonin (20 mg/kg) and zileuton (20 mg/kg) were administered to a unilateral ureteral obstruction (UUO) mouse model, the combination significantly reduced tubular injury and fibrosis by decreasing the expression of profibrotic markers, such as α-SMA and fibronectin. More importantly, the combination ameliorated the increase in 4-HNE levels and decreased GPX4 expression in UUO mice. Overall, the combination of melatonin and zileuton was found to effectively ameliorate ferroptosis-related kidney injury by upregulating the AKT/mTOR/ NRF2 signaling pathway, suggesting a promising therapeutic strategy for protection against ferroptosis-mediated kidney injury and fibrosis.

Keywords

Acknowledgement

This work was supported by a National Research Foundation of Korea (NRF) grant funded by the Korean Government Ministry of Science and Information and Communication Technologies (NRF-2021R1A5A2031612, 2021R1A2B5B03086410, 2022R1G1A1011829, 2022R1A2C1092933) and Inha University Research Grant.

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