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Disulfiram inhibits inflammation and fibrosis in a rat unilateral ureteral obstruction model by inhibiting gasdermin D cleavage and pyroptosis

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Abstract

Background

As an inhibitor of GSDMD, Disulfiram (DSL) can significantly inhibit cell pyroptosis. Cell pyroptosis plays an important role in renal fibrosis.

Methods

HK-2 cells were induced by Lps and ATP to form a pyroptosis model, and the cells were treated by DSL. CCK-8 detected the cell activity. Immunofluorescence (IF) detected the GSDMD. ELISA detected the expression of inflammatory cytokines. Flow cytometry and Western blot detected cell apoptosis and pyroptosis. Collagen type I kit detected collagen secretion, and western blot detected fibrosis marker protein expression. Then, a rat model of unilateral ureteral obstruction (UUO) was established. HE staining detected the degree of renal tissue injury, and Masson staining detected the degree of fibrosis. What’s more, the apoptosis level of tissue cells was detected by TUNEL. And the inflammatory factors in peripheral blood and renal tissue were detected by ELISA. Furthermore, the expression of GSDMD was detected by immunohistochemistry (IHC), and Western blot was used to detect the expression levels of apoptosis and pyroptosis-related proteins in tissues.

Results

It was found that DSL can inhibit the cell membrane perforation of GSDMD-N by inhibiting the cleavage of GSDMD, hence, it inhibited the occurrence of inflammation, cell pyroptosis, and the fibrosis of HK-2 cells. But if the cell has already undergone pyroptosis, DSL does not provide significant prevention. In vivo experiment, it further verified that pretreated DSL had inhibited renal fibrosis injury.

Conclusion

Disulfiram can inhibit inflammation and fibrosis in renal fibrosis rats by inhibiting GSDMD.

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Availability of data and materials

The analyzed data set generated during the present study are available from the corresponding author on reasonable request.

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Author notes

  1. Yu Zhang and Ruicheng Zhang have contribute equally.

Authors and Affiliations

  1. Department of Nephrology, Taizhou People’s Hospital, The Fifth Affiliated Hospital of Nantong University, Taizhou, 225300, Jiangsu, China

    Yu Zhang

  2. Department of Nephrology, Hainan Medical University First Affiliated Hospital, Haikou, 570102, Hainan, China

    Ruicheng Zhang

  3. Department of Blood Purification Center of Nephrology, Nanjing Yimin Hospital, No. 86, Shanggao Road, Jiangning District, Nanjing, 211100, Jiangsu, China

    Xiaohu Han

Authors
  1. Yu Zhang
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  2. Ruicheng Zhang
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Contributions

XH contributed to conception and design, analysis and interpretation of the data critically revised the article for important intellectual content. YZ contributed to the design and analysis of the data, drafted and revised the manuscript. YZ and RZ substantially contributed to the conception and design, acquisition, analysis, and interpretation of data; drafted and critically revised the article for important intellectual content. All authors approved the final version of the article and agreed to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.

Corresponding author

Correspondence to Xiaohu Han.

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Conflict of interest

The authors declare that they have no competing interests.

Ethics approval and consent to participate

The study protocol was approved by ethics committee of Nanjing Yimin Hospital. All of the procedures were in compliance with The National Institutes of Health Guide for the Care and Use of Laboratory Animals.

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Responsible Editor: John Di Battista.

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Zhang, Y., Zhang, R. & Han, X. Disulfiram inhibits inflammation and fibrosis in a rat unilateral ureteral obstruction model by inhibiting gasdermin D cleavage and pyroptosis. Inflamm. Res. 70, 543–552 (2021). https://doi.org/10.1007/s00011-021-01457-y

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  • DOI: https://doi.org/10.1007/s00011-021-01457-y

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