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Pharmacology & ToxicologyActa Biochimica et Biophysica Sinica
Peer-reviewed journal article

Scutellarin inhibits ferroptosis by promoting cellular antioxidant capacity through regulating the Nrf2 signaling

Haiyan YangiD, On-kei Chan, Xiaodi HuangiD, Yan LiangiD, Nuo Sun, Yaping LiiD, Zijian ShiiD, Qingbing ZhaiD, Dong‐Yun OuyangiD, Jinhua LiiD, Xian‐Hui HeiD

Abstract

Ferroptosis is a lytic form of regulated cell death that is driven by iron-dependent lipid peroxidation, and has been implicated in various diseases including acute kidney injury (AKI). Scutellarin is a flavonoid isolated from Erigeron breviscapus (Vant.) Hand.-Mazz. and possesses various pharmacological activities including anti-inflammatory and antioxidative properties. Yet it is unclear whether scutellarin can inhibit ferroptosis and mitigate related diseases. In this study, we found that scutellarin was able to inhibit ferroptosis in both human HK-2 cells and mouse bone marrow-derived macrophages stimulated with RSL3 or erastin. Mitochondrial dysfunction and reactive oxygen species generation were counteracted by scutellarin treatment, suggesting involvement of its antioxidative activity. Furthermore, scutellarin increased the nuclear levels of Nrf2 and the expression of its target genes including HO-1 and GPX4. Scutellarin-mediated inhibition of ferroptosis and an increase of these proteins was abrogated by the co-treatment with brusatol, an Nrf2 inhibitor, indicating an essential role for Nrf2 in this process. In a mouse model of folic acid-induced AKI, scutellarin mitigated acute renal damage as revealed by histopathological analysis and serum blood urea nitrogen and creatinine assays. Folic acid-induced acute renal injury was associated with increased ferroptosis as revealed by elevated levels of 4-hydroxynonenal (4-HNE), a surrogate marker of ferroptosis, which were diminished by scutellarin cotreatment. Specifically, the elevated 4-HNE levels in macrophages (MAC-2 positive) and other renal cells were suppressed by scutellarin. Collectively, scutellarin can inhibit ferroptosis both in cultured cells and in a mouse model of AKI by regulating Nrf2 signaling.

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Claude Sonnet 4.6 · xhigh effort · 8 Nov 2026 · from the full text of version 1 (26 Jun 2025) · CC BY 4.0

Scutellarin suppresses RSL3- and erastin-induced ferroptosis in human HK-2 cells and mouse macrophages via Nrf2 upregulation, and reduces folic acid-induced kidney injury in mice.

Ferroptosis is prominent in acute kidney injury, but it was unknown whether scutellarin, the major ingredient of the clinically used herbal drug breviscapine, can block it. This work tests an Nrf2-driven antioxidant mechanism that differs from commonly used inhibitors such as Fer-1 and liproxstatin-1.

Key results

  1. ShownScutellarin lowered the fraction of PI-positive dying cells in human HK-2 cells and mouse BMDMs challenged with the GPX4 inhibitor RSL3 or the system Xc- inhibitor erastin; ferrostatin-1 blocked the same death. Scutellarin alone was non-toxic.

    HK-2: RSL3 0.4 µM or erastin 10 µM for 5 h; BMDMs: RSL3 10 µM or erastin 10 µM for 24 h; n = 5 fields per condition; ***P < 0.001. Scutellarin at 200 μM or less was non-toxic by WST-1 assay. · Cytotoxicity assay; Scutellarin inhibits ferroptosis both in mouse…; Figure legends

  2. ShownScutellarin reversed RSL3-induced lipid peroxidation signals (C11-BODIPY) and mitochondrial dysfunction (loss of membrane potential by TMRE/JC-1, increased MitoSOX superoxide) in HK-2 cells and BMDMs.

    C11-BODIPY green rose and red fell with RSL3, reversed by scutellarin (200 μM); TMRE, JC-1 and MitoSOX quantified in both cell types; n = 5; **P < 0.01 and ***P < 0.001 (HK-2 RSL3 0.4 µM, BMDM RSL3 10 µM). · Scutellarin inhibits ferroptosis both in mouse…; Scutellarin decreases mitochondrial damage…; Figure legends

  3. ShownScutellarin raised Nrf2, HO-1 and GPX4 protein and nuclear Nrf2 under RSL3 or erastin, and the Nrf2 inhibitor brusatol reversed both the protection from RSL3-induced death and the protein increases in HK-2 cells.

    Immunoblots in HK-2 and BMDMs (n = 3; *P < 0.05, **P < 0.01, ***P < 0.001); nuclear Nrf2 dose-dependently increased (supplementary S2); with brusatol, cell death (n = 5) and Nrf2/GPX4/HO-1 (n = 3) reverted. · Inhibition of ferroptosis by scutellarin relies…; Figure legends

  4. ShownOral scutellarin lessened folic acid-induced acute kidney injury in mice, improving renal histology (less vacuolization and Bowman's space enlargement) and lowering serum BUN and creatinine.

    Female C57BL/6J mice, n = 5 per group in four groups; scutellarin 200 mg/kg by gavage, folic acid 250 mg/kg i.p., sampled 24 h later; BUN and creatinine significantly decreased vs folic acid alone (**P < 0.01, ***P < 0.001). · Animal model of AKI; Scutellarin administration mitigates folic…; Figure legends

  5. SuggestedIn injured kidneys, scutellarin was associated with higher GPX4, Nrf2 and HO-1 and lower 4-HNE protein adducts; 4-HNE staining in MAC-2-positive macrophages and other renal cells was reduced.

    Folic acid lowered GPX4 and raised 4-HNE-modified proteins; scutellarin reversed this. Western blot used kidneys from 3 mice per group (n = 3; *P < 0.05, **P < 0.01, ***P < 0.001); 4-HNE by immunofluorescence. · Mitigation of acute kidney injury by scutellarin…; Figure legends

Methods, data and reproducibility

HK-2 cells and mouse BMDMs were challenged with RSL3 or erastin and read out by PI/Hoechst death staining, fluorescent probes for lipid peroxidation and mitochondria, and immunoblotting, with brusatol to inhibit Nrf2. A folic acid AKI mouse model was assessed by histology, serum BUN/creatinine, immunoblotting and 4-HNE immunofluorescence.

  • HK-2 human renal proximal tubular epithelial cell line, treated with RSL3 or erastin, scutellarin and brusatol · human · n = n = 5 fields for death assays; n = 3 for immunoblots
  • Mouse bone marrow-derived macrophages (BMDMs) treated with RSL3 or erastin and scutellarin · mouse · n = n = 5 for death and imaging assays; n = 3 for immunoblots
  • Folic acid-induced AKI in female C57BL/6J mice (6-8 weeks): vehicle, scutellarin, folic acid, scutellarin + folic acid · mouse · n = n = 5 per group; kidneys from 3 mice per group for western blot
Methods & tools
Scutellarin, RSL3, Erastin, Ferrostatin-1, Brusatol, PI and Hoechst 33342 staining, C11-BODIPY 581/591, MitoSOX Red, JC-1 and TMRE, WST-1 assay, ImageJ, GraphPad Prism 7.0
Reagents & environment
Statistics in GraphPad Prism 7.0; densitometry with ImageJ and a FluorChem 8000 system; imaging on Zeiss Axio Observer D1 and ZEN software.
Reproducibility gaps
No code or data repository is given. Raw images and blots are not deposited, and supplementary Figures S1 and S2 are cited but not included in the provided text.
Limitations (5)
  • The authors lack direct evidence that the kidney was effectively exposed to orally given scutellarin. They rely on earlier pharmacokinetic studies indicating kidney distribution despite low bioavailability, so repeated dosing may be needed. · stated by the authors
  • How scutellarin protects mitochondria is unresolved; possible routes (Nrf2 targets, PDK-PDC axis, direct radical trapping) are untested, and effects on system Xc- are unknown. · stated by the authors
  • Other mechanisms may act in parallel with Nrf2 signaling in vivo. · stated by the authors
  • Nrf2 dependence rests on the pharmacological inhibitor brusatol in HK-2 cells with RSL3; no genetic Nrf2 loss-of-function or in vivo Nrf2 inhibition is reported, and brusatol may have other effects. · noted in this summary
  • The in vivo evidence comes from a single folic acid model with one scutellarin dose, small groups, and immunoblots from 3 mice per group; no ferroptosis inhibitor positive control is included. · noted in this summary

Who should read:Read this if you study natural-product ferroptosis inhibitors, Nrf2-GPX4 antioxidant signaling, or ferroptosis in acute kidney injury models.

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