Mechanism of Circadian Regulation in Ferroptosis of the BMAL1/NRF2 Pathway in Renal Ischemia–Reperfusion

<b>Background</b>: Renal ischemia–reperfusion injury (IRI) is a frequent cause of kidney transplant failure. Recent studies have shown that the extent of injury is closely linked to ferroptosis, and the process of cellular ferroptosis is diurnal and regulated by circadian genes. <i>...

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Main Authors: Shang Xu, Qiao Tang, Haiyang Du, Jiatao Xie, Ruoxin He, Ruiyan Wang, Qian Sun
Format: Article
Language:English
Published: MDPI AG 2025-06-01
Series:Biomedicines
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Online Access:https://www.mdpi.com/2227-9059/13/6/1375
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author Shang Xu
Qiao Tang
Haiyang Du
Jiatao Xie
Ruoxin He
Ruiyan Wang
Qian Sun
author_facet Shang Xu
Qiao Tang
Haiyang Du
Jiatao Xie
Ruoxin He
Ruiyan Wang
Qian Sun
author_sort Shang Xu
collection DOAJ
description <b>Background</b>: Renal ischemia–reperfusion injury (IRI) is a frequent cause of kidney transplant failure. Recent studies have shown that the extent of injury is closely linked to ferroptosis, and the process of cellular ferroptosis is diurnal and regulated by circadian genes. <i>NRF2</i>, involved in iron–heme metabolism, may be related to ferroptosis. We hypothesize that the pathway plays a role in circadian regulation in ferroptosis in renal IRI. <b>Methods</b>: Using hematoxylin and eosin (H&E) staining, enzyme-linked immunosorbent assay (ELISA), Cell Counting Kit-8 (CCK8), flow cytometry, real-time quantitative reverse transcription PCR (qRT-PCR), and Western blotting, we analyzed renal tubular tissues in vivo and in vitro and compared the groups with IR injury treatment, inhibition of ferroptosis, and inhibition of <i>BMAL1</i> gene expression at the ZT0 (zeitgeber time 0) and ZT12 (zeitgeber time 12) time points. <b>Results</b>: IR injury treatments caused exacerbation of injury, both in vivo and in vitro, and were more pronounced at the ZT12 time point, which correlates with circadian rhythms. The use of the ferroptosis inhibitor (Fer-I) attenuated IR injury, suggesting that IRI is associated with ferroptosis. In contrast, reduced <i>BMAL1</i>-gene expression exacerbated injury, and <i>NRF2</i>, which is elevated in IR injury, was suppressed. <b>Conclusions</b>: The circadian gene <i>BMAL1</i> affects the circadian rhythm of ferroptosis in renal IRI through the regulation of <i>NRF2</i> and its downstream pathway. In this study, renal injury is well ameliorated by the ferroptosis inhibitor, exhibiting potential as a therapeutic agent for use in renal transplantation.
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spelling doaj-art-94fa2d928c0e46b5881aa7cc61eef9d62025-08-20T03:27:26ZengMDPI AGBiomedicines2227-90592025-06-01136137510.3390/biomedicines13061375Mechanism of Circadian Regulation in Ferroptosis of the BMAL1/NRF2 Pathway in Renal Ischemia–ReperfusionShang Xu0Qiao Tang1Haiyang Du2Jiatao Xie3Ruoxin He4Ruiyan Wang5Qian Sun6Department of Anesthesiology, Renmin Hospital of Wuhan University, No. 238 Jiefang Road, Wuchang District, Wuhan 430061, ChinaDepartment of Anesthesiology, Renmin Hospital of Wuhan University, No. 238 Jiefang Road, Wuchang District, Wuhan 430061, ChinaDepartment of Anesthesiology, Renmin Hospital of Wuhan University, No. 238 Jiefang Road, Wuchang District, Wuhan 430061, ChinaDepartment of Anesthesiology, Renmin Hospital of Wuhan University, No. 238 Jiefang Road, Wuchang District, Wuhan 430061, ChinaDepartment of Anesthesiology, Renmin Hospital of Wuhan University, No. 238 Jiefang Road, Wuchang District, Wuhan 430061, ChinaDepartment of Anesthesiology, Renmin Hospital of Wuhan University, No. 238 Jiefang Road, Wuchang District, Wuhan 430061, ChinaDepartment of Anesthesiology, Renmin Hospital of Wuhan University, No. 238 Jiefang Road, Wuchang District, Wuhan 430061, China<b>Background</b>: Renal ischemia–reperfusion injury (IRI) is a frequent cause of kidney transplant failure. Recent studies have shown that the extent of injury is closely linked to ferroptosis, and the process of cellular ferroptosis is diurnal and regulated by circadian genes. <i>NRF2</i>, involved in iron–heme metabolism, may be related to ferroptosis. We hypothesize that the pathway plays a role in circadian regulation in ferroptosis in renal IRI. <b>Methods</b>: Using hematoxylin and eosin (H&E) staining, enzyme-linked immunosorbent assay (ELISA), Cell Counting Kit-8 (CCK8), flow cytometry, real-time quantitative reverse transcription PCR (qRT-PCR), and Western blotting, we analyzed renal tubular tissues in vivo and in vitro and compared the groups with IR injury treatment, inhibition of ferroptosis, and inhibition of <i>BMAL1</i> gene expression at the ZT0 (zeitgeber time 0) and ZT12 (zeitgeber time 12) time points. <b>Results</b>: IR injury treatments caused exacerbation of injury, both in vivo and in vitro, and were more pronounced at the ZT12 time point, which correlates with circadian rhythms. The use of the ferroptosis inhibitor (Fer-I) attenuated IR injury, suggesting that IRI is associated with ferroptosis. In contrast, reduced <i>BMAL1</i>-gene expression exacerbated injury, and <i>NRF2</i>, which is elevated in IR injury, was suppressed. <b>Conclusions</b>: The circadian gene <i>BMAL1</i> affects the circadian rhythm of ferroptosis in renal IRI through the regulation of <i>NRF2</i> and its downstream pathway. In this study, renal injury is well ameliorated by the ferroptosis inhibitor, exhibiting potential as a therapeutic agent for use in renal transplantation.https://www.mdpi.com/2227-9059/13/6/1375ferroptosisrenal ischemia–reperfusion injurycircadian rhythm<i>BMAL1</i><i>NRF2</i>
spellingShingle Shang Xu
Qiao Tang
Haiyang Du
Jiatao Xie
Ruoxin He
Ruiyan Wang
Qian Sun
Mechanism of Circadian Regulation in Ferroptosis of the BMAL1/NRF2 Pathway in Renal Ischemia–Reperfusion
Biomedicines
ferroptosis
renal ischemia–reperfusion injury
circadian rhythm
<i>BMAL1</i>
<i>NRF2</i>
title Mechanism of Circadian Regulation in Ferroptosis of the BMAL1/NRF2 Pathway in Renal Ischemia–Reperfusion
title_full Mechanism of Circadian Regulation in Ferroptosis of the BMAL1/NRF2 Pathway in Renal Ischemia–Reperfusion
title_fullStr Mechanism of Circadian Regulation in Ferroptosis of the BMAL1/NRF2 Pathway in Renal Ischemia–Reperfusion
title_full_unstemmed Mechanism of Circadian Regulation in Ferroptosis of the BMAL1/NRF2 Pathway in Renal Ischemia–Reperfusion
title_short Mechanism of Circadian Regulation in Ferroptosis of the BMAL1/NRF2 Pathway in Renal Ischemia–Reperfusion
title_sort mechanism of circadian regulation in ferroptosis of the bmal1 nrf2 pathway in renal ischemia reperfusion
topic ferroptosis
renal ischemia–reperfusion injury
circadian rhythm
<i>BMAL1</i>
<i>NRF2</i>
url https://www.mdpi.com/2227-9059/13/6/1375
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