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Ferroptosis in the colon epithelial cells as a therapeutic target for ulcerative colitis

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A Letter to the Editor to this article was published on 23 November 2023

Abstract

Background

Ferroptosis, a type of programmed cell death triggered by oxidative stress, was suspected to play a role in ulcerative colitis. Indigo naturalis is highly effective against ulcerative colitis, but its mechanism is unclear. This study found that indigo naturalis treatment suppressed ferroptosis.

Methods

We analyzed 770 mRNA expressions of patients with ulcerative colitis. Suppression of ferroptosis by indigo naturalis treatment was shown using a cell death assay. Malondialdehyde levels and reactive oxygen species were analyzed in CaCo-2 cells treated with indigo naturalis. Glutathione metabolism was shown by metabolomic analysis. Extraction of the ingredients indigo naturalis from the rectal mucosa was performed using liquid chromatograph—mass spectrometry.

Results

Gene expression profiling showed that indigo naturalis treatment increased antioxidant genes in the mucosa of patients with ulcerative colitis. In vitro analysis showed that nuclear factor erythroid-2-related factor 2-related antioxidant gene expression was upregulated by indigo naturalis. Indigo naturalis treatment rendered cells resistant to ferroptosis. Metabolomic analysis suggested that an increase in reduced glutathione by indigo naturalis. The protein expression of CYP1A1 and GPX4 was increased in the rectum by treatment with indigo naturalis. The main ingredients of indigo naturalis, indirubin and indigo inhibited ferroptosis. Indirubin was detected in the rectal mucosa of patients with ulcerative colitis who were treated with indigo naturalis.

Conclusions

Suppression of ferroptosis by indigo naturalis in the intestinal epithelium could be therapeutic target for ulcerative colitis. The main active ingredient of indigo naturalis may be indirubin.

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Abbreviations

AHR:

Aryl hydrocarbon receptor

CYP1A1:

Cytochrome P450 1a1

Cys:

Cysteine

DMSO:

Dimethyl sulfoxide

FICZ:

6-Formylindolo[3, 2-b]carbazole

GABA:

Gamma-aminobutyric acid

γ-Glu-Cys:

Gamma-glutamylcysteine

GCL:

Glutamate-cysteine ligase

GCLC:

Glutamate-cysteine ligase catalytic subunit

GCLM:

Glutamate-cysteine ligase modifier subunit

GPX4:

Glutathione peroxidase 4

GSH:

Reduced glutathione

GSSG:

Glutathione disulfide

GSTP1:

Pi-class glutathione S-transferase

Gln:

Glutamine

Glu:

Glutamic acid

Gly:

Glycine

HDAC2:

Histone deacetylase 2

H2O2 :

Hydrogen peroxide

HO-1:

Heme oxygenase-1

IN:

Indigo naturalis

LC/MS:

Liquid chromatography—mass spectrometry

MDA:

Malondialdehyde

NADPH:

Nicotinamide adenine dinucleotide phosphate

NRF2:

Nuclear factor erythroid-2-related factor 2

PRDX2:

Peroxiredoxin 2

ROS:

Reactive oxygen species

PCR:

Polymerase chain reaction

SELENBP1:

Selenium binding protein 1

SOD1:

Superoxide dismutase 1

SOD2:

Superoxide dismutase 2

SRXN:

Sulfiredoxin-1

Ser:

Serine

TCDD:

Tetrachlorodibenzo-para-dioxin

TNF:

Tumor necrosis factor

TXN:

Thioredoxin 1

TXNR:

Thioredoxin reductase 1

UC:

Ulcerative colitis

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Acknowledgements

We thank Ellen Knapp, PhD, from Edanz (https://jp.edanz.com/ac) for editing a draft of this manuscript. We appreciate the technical assistance from The Research Support Center, Research Center for Human Disease Modeling, Kyushu University Graduate School of Medical Sciences.

Funding

This work was supported by JSPS KAKENHI grants ‘[Grant-in-Aid for Scientific Research (C) JP21K06783 and Grant-in-Aid for Young Scientists (B) JP22K15668]’.

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Authors and Affiliations

  1. Department of Medicine and Clinical Science, Graduate School of Medical Science, Kyushu University, Maidashi 3-1-1, Higashi-ku, Fukuoka, 812-8582, Japan

    Akihito Yokote, Noriyuki Imazu, Junji Umeno, Keisuke Kawasaki, Yuta Fuyuno, Yuichi Matsuno, Takanari Kitazono & Takehiro Torisu

  2. Department of Endoscopic Diagnostics and Therapeutics, Kyushu University Hospital, Fukuoka, 812-8582, Japan

    Shin Fujioka

  3. International Medical Department, Kyushu University Hospital, Fukuoka, 812-8582, Japan

    Tomohiko Moriyama

  4. Department of Medicine and Biosystemic Science, Graduate School of Medical Sciences, Kyushu University, Fukuoka, 812-8582, Japan

    Kohta Miyawaki & Koichi Akashi

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  1. Akihito Yokote
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Contributions

AY, NI and TT conceived the idea of the study. AY, and TT were involved in the data analysis. JU, KK, SF, YF, YM, TM, and TT collected the clinical data and contributed to data interpretation. AYo, KM, KA, TK, and TT contributed to drafting of the manuscript and critical revision. All authors revised the manuscript, approved the manuscript to be published, and agree to be accountable for all aspects of the work.

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Correspondence to Takehiro Torisu.

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Yokote, A., Imazu, N., Umeno, J. et al. Ferroptosis in the colon epithelial cells as a therapeutic target for ulcerative colitis. J Gastroenterol 58, 868–882 (2023). https://doi.org/10.1007/s00535-023-02016-4

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