Rosmarinic acid plus deferasirox inhibits ferroptosis to alleviate crush syndrome-related AKI via Nrf2/Keap1 pathway

Ou Qiao; Li Zhang; Lu Han; Xinyue Wang; Zizheng Li; Fengjiao Bao; Herui Hao; Yingjie Hou; Xiaohong Duan; Ning Li; Yanhua Gong

doi:10.1016/j.phymed.2024.155700

Rosmarinic acid plus deferasirox inhibits ferroptosis to alleviate crush syndrome-related AKI via Nrf2/Keap1 pathway

Phytomedicine. 2024 May 1:129:155700. doi: 10.1016/j.phymed.2024.155700. Online ahead of print.

Authors

Ou Qiao¹, Li Zhang¹, Lu Han¹, Xinyue Wang¹, Zizheng Li¹, Fengjiao Bao¹, Herui Hao¹, Yingjie Hou¹, Xiaohong Duan¹, Ning Li², Yanhua Gong³

Affiliations

¹ Medical School, Faculty of Medicine, Tianjin University, No. 92 Weijin Road, Nankai District, Tianjin, 300072, China; Institute of Disaster and Emergency Medicine, Faculty of Medicine, Tianjin University, China.
² Medical School, Faculty of Medicine, Tianjin University, No. 92 Weijin Road, Nankai District, Tianjin, 300072, China; Institute of Disaster and Emergency Medicine, Faculty of Medicine, Tianjin University, China; Key Laboratory for Disaster Medicine Technology, Tianjin, China. Electronic address: lining620@tju.edu.cn.
³ Medical School, Faculty of Medicine, Tianjin University, No. 92 Weijin Road, Nankai District, Tianjin, 300072, China; Institute of Disaster and Emergency Medicine, Faculty of Medicine, Tianjin University, China; Key Laboratory for Disaster Medicine Technology, Tianjin, China. Electronic address: gongyanhua@tju.edu.cn.

PMID: 38704914
DOI: 10.1016/j.phymed.2024.155700

Abstract

Background: Myoglobin (Mb) induced death of renal tubular epithelial cells (RTECs) is a major pathological factor in crush syndrome-related acute kidney injury (CS-AKI). It is unclear whether ferroptosis is involved and could be a target for treatment.

Purpose: This study aimed to evaluate the potential therapeutic effects of combining the natural small molecule rosemarinic acid (RA) and the iron chelator deferasirox (Dfe) on CS-AKI through inhibition of ferroptosis.

Methods: Sequencing data were downloaded from the GEO database, and differential expression analysis was performed using the R software limma package. The CS-AKI mouse model was constructed by squeezing the bilateral thighs of mice for 16 h with 1.5 kg weight. TCMK1 and NRK-52E cells were induced with 200 μM Mb and then treated with RA combined with Dfe (Dfe + RA, both were 10 μM). Functional and pathological changes in mouse kidney were evaluated by glomerular filtration rate (GFR) and HE pathology. Immunofluorescence assay was used to detect Mb levels in kidney tissues. The expression levels of ACSL4, GPX4, Keap1, and Nrf2 were analyzed by WB.

Results: We found that AKI mice in the GSE44925 cohort highly expressed the ferroptosis markers ACSL4 and PTGS2. CS-AKI mice showed a rapid decrease in GFR, up-regulation of ACSL4 expression in kidney tissue, and down-regulation of GPX4 expression, indicating activation of the ferroptosis pathway. Mb was found to deposit in renal tubules, and it has been proven to cause ferroptosis in TCMK1 and NRK-52E cells in vitro. We found that Dfe had a strong iron ion scavenging effect and inhibited ACSL4 expression. RA could disrupt the interaction between Keap1 andNrf2, stabilize Nrf2, and promote its nuclear translocation, thereby exerting antioxidant effects. The combination of Dfe and RA effectively reversed Mb induced ferroptosis in RTECs.

Conclusion: In conclusion, we found that RA combined with Dfe attenuated CS-AKI by inhibiting Mb-induced ferroptosis in RTECs via activating the Nrf2/Keap1 pathway.

Keywords: AKI; Crush syndrome; Deferasirox; Ferroptosis; Myoglobin; Rosemarinic acid.