CoChat
Sign Up
Cited by 316Selenium in Biological SystemsAntioxidant Activity and Oxidative StressRedox biology and oxidative stress

The DOI matches its Crossref record; 39 of the 40 references it cites were found in a registry or an index.

Glutathione peroxidase 4 and vitamin E cooperatively prevent hepatocellular degeneration

Open access
Carlson, Bradley A., Tobe, Ryuta, Yefremova, Elena, Tsuji, Petra A., Hoffmann, Victoria J., Schweizer, Ulrich, Gladyshev, Vadim N., Hatfield, Dolph L., Conrad, Marcus·2016·Redox Biology
DOI 10.1016/j.redox.2016.05.003PMC PMC4900515
Open this paper in CoChat — free accountOpen PDF

Abstract

The selenoenzyme glutathione peroxidase 4 (Gpx4) is an essential mammalian glutathione peroxidase, which protects cells against detrimental lipid peroxidation and governs a novel form of regulated necrotic cell death, called ferroptosis. To study the relevance of Gpx4 and of another vitally important selenoprotein, cytosolic thioredoxin reductase (Txnrd1), for liver function, mice with conditional deletion of Gpx4 in hepatocytes were studied, along with those lacking Txnrd1 and selenocysteine (Sec) tRNA ( Trsp ) in hepatocytes. Unlike Txnrd1 - and Trsp -deficient mice, Gpx4 −/− mice died shortly after birth and presented extensive hepatocyte degeneration. Similar to Txnrd1 -deficient livers, Gpx4 −/− livers manifested upregulation of nuclear factor (erythroid-derived)-like 2 (Nrf2) response genes. Remarkably, Gpx4 −/− pups born from mothers fed a vitamin E-enriched diet survived, yet this protection was reversible as subsequent vitamin E deprivation caused death of Gpx4 -deficient mice ~4 weeks thereafter. Abrogation of selenoprotein expression in Gpx4 −/− mice did not result in viable mice, indicating that the combined deficiency aggravated the loss of Gpx4 in liver. By contrast, combined Trsp / Txnrd1 -deficient mice were born, but had significantly shorter lifespans than either single knockout, suggesting that Txnrd1 plays an important role in supporting liver function of mice lacking Trsp . In sum our study demonstrates that the ferroptosis regulator Gpx4 is critical for hepatocyte survival and proper liver function, and that vitamin E can compensate for its loss by protecting cells against deleterious lipid peroxidation.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1016/j.redox.2016.05.003

Cites

39 of 39 references verified0 citation errors · 0 not checkable

Statistics

Figures

Graphical abstract
Graphical abstract CC BY-NC-ND · PubMed Central
Fig. 1 Liver necrosis in Alb-cre; Gpx4fl/fl mice. H&E staining of Gpx4 control and Alb-cre; Gpx4fl/fl livers at 10× and 40× magnification from 1 day old pups revealed extensive hepatocyte degeneration and necrosis, lack of veins (arrow) and sinusoids in livers of Gpx4-deficient pups.
Fig. 1 Liver necrosis in Alb-cre; Gpx4fl/fl mice. H&E staining of Gpx4 control and Alb-cre; Gpx4fl/fl livers at 10× and 40× magnification from 1 day old pups revealed extensive hepatocyte degeneration and necrosis, lack of veins (arrow) and sinusoids in livers of Gpx4-deficient pups.CC BY-NC-ND · PubMed Central
Fig. 2 Expression levels of selenoproteins in livers of 1 day old control, Trsp-, Txnrd1- and Gpx4-deficient mice. (A) mRNA levels of Gpx1, Gpx2, Gpx4, Txnrd1, Selr, and Sepw were analyzed by qPCR. Data are shown as relative mRNA levels normalized to Gapdh in all liver samples (n=3 for each genotype). *Denotes statistical difference (P<0.05). (B) Protein levels of Gpx1, Gpx4, Txnrd1, SelR and SepW were analyzed by immunoblotting in both control and knockout liver samples (n=2 each). Gapdh levels are used as a control for protein loading. (C) Txnrd1 activity in livers of control, Trsp-, Txnrd1- and Gpx4-deficient mice. Data are the mean±SEM for three independent experiments and are expressed as µmol TNB/min/mg protein. *Denotes statistical difference (P<0.05).
Fig. 2 Expression levels of selenoproteins in livers of 1 day old control, Trsp-, Txnrd1- and Gpx4-deficient mice. (A) mRNA levels of Gpx1, Gpx2, Gpx4, Txnrd1, Selr, and Sepw were analyzed by qPCR. Data are shown as relative mRNA levels normalized to Gapdh in all liver samples (n=3 for each genotype). *Denotes statistical difference (P<0.05). (B) Protein levels of Gpx1, Gpx4, Txnrd1, SelR and SepW were analyzed by immunoblotting in both control and knockout liver samples (n=2 each). Gapdh levels are used as a control for protein loading. (C) Txnrd1 activity in livers of control, Trsp-, Txnrd1- and Gpx4-deficient mice. Data are the mean±SEM for three independent experiments and are expressed as µmol TNB/min/mg protein. *Denotes statistical difference (P<0.05).CC BY-NC-ND · PubMed Central
Fig. 3 mRNA expression levels of Nrf2-regulated genes in livers of 1 day old control, Trsp-, Txnrd1- and Gpx4-deficient mice. (A) mRNA levels of Gsta1, Gclc, Gsr, Srxn1 and Cbr3 were analyzed by qPCR. Data are shown as relative mRNA levels normalized to Gapdh in all liver samples (n=3 for each genotype). *Denotes statistical difference (P<0.05). (B) Total GSH was measured spectrophotometrically and the levels calculated relative to the levels found in control mice. Total GSH concentrations are the mean±S. D. for three independent experiments.
Fig. 3 mRNA expression levels of Nrf2-regulated genes in livers of 1 day old control, Trsp-, Txnrd1- and Gpx4-deficient mice. (A) mRNA levels of Gsta1, Gclc, Gsr, Srxn1 and Cbr3 were analyzed by qPCR. Data are shown as relative mRNA levels normalized to Gapdh in all liver samples (n=3 for each genotype). *Denotes statistical difference (P<0.05). (B) Total GSH was measured spectrophotometrically and the levels calculated relative to the levels found in control mice. Total GSH concentrations are the mean±S. D. for three independent experiments.CC BY-NC-ND · PubMed Central
Fig. 4 Survival rate of Gpx4 control and Alb-cre; Gpx4fl/fl mice following removal of vitamin E from the diet. (A) Schematic representation of experimental design and feeding plan of mice maintained on vitamin E-deficient (−vitamin E) or vitamin E-supplemented (+vitamin E) diets. (B) Survival curve of control (n=12) and Alb-cre; Gpx4fl/fl (n=10) mice on defined diets. Arrow notes the time when mice were placed on a vitamin E-deficient diet. A log-rank test was used for statistical analysis. *Denotes statistical difference (P<0.0001).
Fig. 4 Survival rate of Gpx4 control and Alb-cre; Gpx4fl/fl mice following removal of vitamin E from the diet. (A) Schematic representation of experimental design and feeding plan of mice maintained on vitamin E-deficient (−vitamin E) or vitamin E-supplemented (+vitamin E) diets. (B) Survival curve of control (n=12) and Alb-cre; Gpx4fl/fl (n=10) mice on defined diets. Arrow notes the time when mice were placed on a vitamin E-deficient diet. A log-rank test was used for statistical analysis. *Denotes statistical difference (P<0.0001).CC BY-NC-ND · PubMed Central
Fig. 5 Liver necrosis in Alb-cre; Gpx4fl/fl mice following removal of vitamin E. (A) H&E staining of Gpx4 control and two Alb-cre; Gpx4fl/fl livers at 10× and 20× magnification from mice that have been maintained on a vitamin E-sufficient diet for 6 weeks, followed by removal of vitamin E for 3 weeks. N=areas of necrosis. Arrows point to areas of acute inflammation. (B) TBARS assay was used to assess plasma levels of malondialdehyde (MDA) in Gpx4 control (n=3) and Alb-cre; Gpx4fl/fl (n=3) mice maintained on a vitamin E-supplemented diet (vitamin E) for nine weeks and in Gpx4 control (n=3) and Alb-cre; Gpx4fl/fl (n=3) mice maintained on a vitamin E-supplemented diet for six weeks followed by removal of vitamin E (- vitamin E) for 3 weeks. *Denotes statistical difference (P<0.05).
Fig. 5 Liver necrosis in Alb-cre; Gpx4fl/fl mice following removal of vitamin E. (A) H&E staining of Gpx4 control and two Alb-cre; Gpx4fl/fl livers at 10× and 20× magnification from mice that have been maintained on a vitamin E-sufficient diet for 6 weeks, followed by removal of vitamin E for 3 weeks. N=areas of necrosis. Arrows point to areas of acute inflammation. (B) TBARS assay was used to assess plasma levels of malondialdehyde (MDA) in Gpx4 control (n=3) and Alb-cre; Gpx4fl/fl (n=3) mice maintained on a vitamin E-supplemented diet (vitamin E) for nine weeks and in Gpx4 control (n=3) and Alb-cre; Gpx4fl/fl (n=3) mice maintained on a vitamin E-supplemented diet for six weeks followed by removal of vitamin E (- vitamin E) for 3 weeks. *Denotes statistical difference (P<0.05).CC BY-NC-ND · PubMed Central
Fig. 6 Survival curve of combined Trsp- and Txnrd1-deficient mice. Survival curve of control (n=10), Alb-cre: Trspfl/fl; Txnrd1fl/fl(n=6), Alb-cre: Trspfl/fl; Txnrd1fl/+ (n=6) and Alb-cre: Trspfl/fl; Txnrd1+/+ mice (n=10) maintained on standard chow. A log-rank test was used for all statistical analyses. *Denotes statistical difference (P<0.0001) from control.
Fig. 6 Survival curve of combined Trsp- and Txnrd1-deficient mice. Survival curve of control (n=10), Alb-cre: Trspfl/fl; Txnrd1fl/fl(n=6), Alb-cre: Trspfl/fl; Txnrd1fl/+ (n=6) and Alb-cre: Trspfl/fl; Txnrd1+/+ mice (n=10) maintained on standard chow. A log-rank test was used for all statistical analyses. *Denotes statistical difference (P<0.0001) from control.CC BY-NC-ND · PubMed Central

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Cited by

Sorted by citations

See all 316

Related

See all

Metadata

Authors
Carlson, B. A., Tobe, R., Yefremova, E., Tsuji, P. A., Hoffmann, V. J., Schweizer, U., Gladyshev, V. N., Hatfield, D. L., & Conrad, M.
Affiliations
  • National Institutes of Health (US)
  • Helmholtz Munich (DE)
  • Towson University (US)
  • University of Bonn (DE)
  • Brigham and Women's Hospital (US)
Corresponding author
  • Hatfield, Dolph L.
  • Conrad, Marcus
ORCID
Venue
Redox Biology
Publisher
Elsevier BV
Published
2016-10
Volume
9
Pages
22-31
ISSN
2213-2317 (ISSN-L)
DOI
10.1016/j.redox.2016.05.003
PMC
PMC4900515
OpenAlex
W2408722709
Cited by
316
References
40
Citations by year
2026: 27 · 2025: 38 · 2024: 52 · 2023: 40 · 2022: 47 · 2021: 27 · 2020: 22 · 2019: 16 · 2018: 24 · 2017: 14 · 2016: 8
FWCI
21.64
Type
journal article
Language
en
Open access
Licence
Locations
  • Redox Biology (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • PuSH - Publication Server of Helmholtz Zentrum München (submittedVersion, cc-by)
  • Digital Access to Scholarship at Harvard (DASH) (Harvard University) (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Site cant be reached (submittedVersion, cc-by)
Funders
  • National Institutes of Health
  • National Cancer Institute
  • Center for Cancer Research
  • Deutsche Forschungsgemeinschaft (CO 291/2-3, CO 291/5-1)
  • Human Frontier Science Program (RGP0013/14)
Topics
  • Health Sciences › Nursing › Nutrition and Dietetics › Selenium in Biological Systems
  • Health Sciences › Medicine › Biochemistry (medical) › Antioxidant Activity and Oxidative Stress
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
Keywords
GPX4, vitamin E, ferroptosis, hepatocyte degeneration, selenoproteins, thioredoxin reductase 1, selenocysteine tRNA, lipid peroxidation, conditional knockout mice, liver function, hepatocyte viability
MeSH terms
Phospholipid Hydroperoxide Glutathione Peroxidase, Animals, Cell Survival, Female, Gene Expression Regulation, Genotype, Glutathione, Glutathione Peroxidase, Liver, Liver Regeneration, Male, Necrosis, Vitamin E, Lipid Peroxidation, Mice, Knockout, Glutathione Disulfide, Hepatocytes, NF-E2-Related Factor 2, Mice

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review