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CircDCBLD2 alleviates liver fibrosis by regulating ferroptosis via facilitating STUB1-mediated PARK7 ubiquitination degradation

  • Original Article―Liver, Pancreas, and Biliary Tract
  • Published:
Journal of Gastroenterology Aims and scope Submit manuscript

Abstract

Background

Liver fibrosis can progress to cirrhosis and hepatic carcinoma without treatment. CircDCBLD2 was found to be downregulated in liver fibrosis. However, the precise underlying mechanism requires further investigation.

Methods

qRT-PCR, Western blot, and immunohistochemistry assays were used to detect the related molecule levels. HE, Masson’s trichrome, and Sirius Red staining were used to assess the pathological changes in mice’s liver tissues. Flow cytometric analysis and commercial kit were used to assess the levels of lipid reactive oxygen species (ROS), malonaldehyde (MDA), glutathione (GSH), and iron. Cell viability was assessed by MTT. Immunoprecipitation was used to study the ubiquitination of PARK7. Mitophagy was determined by immunostaining and confocal imaging. RIP and Co-IP assays were used to assess the interactions of circDCBLD2/HuR, HuR/STUB1, and STUB1/PARK7. Fluorescence in situ hybridization and immunofluorescence staining were used to assess the co-localization of circDCBLD2 and HuR.

Results

CircDCBLD2 was downregulated, whereas PARK7 was upregulated in liver fibrosis. Ferroptosis activators increased circDCBLD2 while decreasing PARK7 in hepatic stellate cells (HSCs) and mice with liver fibrosis. CircDCBLD2 overexpression reduced cell viability and GSH, PARK7, and GPX4 expression in erastin-treated HSCs while increasing MDA and iron levels, whereas circDCBLD2 knockdown had the opposite effect. CircDCBLD2 overexpression increased STUB1-mediated PARK7 ubiquitination by promoting HuR-STUB1 binding and thus increasing STUB1 mRNA stability. PARK7 overexpression or HuR knockdown reversed the effects of circDCBLD2 overexpression on HSC activation and ferroptosis. CircDCBLD2 reduced liver fibrosis in mice by inhibiting PARK7.

Conclusion

CircDCBLD2 overexpression increased PARK7 ubiquitination degradation by upregulating STUB1 through its interaction with HuR, inhibiting HSC activation and promoting HSC ferroptosis, ultimately enhancing liver fibrosis.

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Abbreviations

HSCs:

Hepatic stellate cells

GSH:

Glutathione

GPX4:

Glutathione peroxidase 4

ROS:

Reactive oxygen species

PARK7:

Parkinson disease 7

STUB1:

STIP1 homology and U-Box containing protein 1

circRNAs:

Circular RNAs

FBS:

Fetal bovine serum

shRNA:

Short hairpin RNA

act-D:

Actinomycin D

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide

MDA:

Malondialdehyde

RIP:

RNA immunoprecipitation

CHX:

Cycloheximide

Co-IP:

Co-immunoprecipitation

CCl4 :

Carbon tetrachloride

H&E:

Hematoxylin and eosin

IHC:

Immunohistochemistry

ALT:

Alanine aminotransferase

AST:

Aspartate aminotransferase

HYP:

Hydroxyproline

qRT-PCR:

Quantitative real-time PCR

PVDF:

Polyvinylidene difluoride

HO-1:

Heme oxygenase-1

IRP2:

Iron-regulatory protein 2

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Funding

This study was funded by grants from the National Natural Science Foundation of China (No. 82170639), the National Natural Science Foundation of China Youth Fund (No. 82200671), and the Hunan Provincial Natural Science Foundation (No. 2022JJ70068, 2022JJ70070).

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Authors and Affiliations

  1. Department of Infectious Disease, Third Xiangya Hospital, Central South University, Hunan, 410013, China

    Juan Wang, Haoye Zhang, Limin Chen, Kangkang Fu, Yu Yan & Zhenguo Liu

  2. Changsha & Hunan Key Laboratory of Viral Hepatitis, Xiangya Hospital, Central South University, Hunan, 410008, China

    Zhenguo Liu

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  1. Juan Wang
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Contributions

JW: conceptualization; methodology; HZ: visualization; supervision; LC: validation; formal analysis; KF: investigation; resources; YY: data curation; writing–original draft; ZL: writing—review and editing; project administration; funding acquisition.

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Correspondence to Zhenguo Liu.

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The Animal Care and Use Committee of Central South University’s Third Xiangya Hospital approved all mouse-related experiments.

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Wang, J., Zhang, H., Chen, L. et al. CircDCBLD2 alleviates liver fibrosis by regulating ferroptosis via facilitating STUB1-mediated PARK7 ubiquitination degradation. J Gastroenterol 59, 229–249 (2024). https://doi.org/10.1007/s00535-023-02068-6

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  • DOI: https://doi.org/10.1007/s00535-023-02068-6

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