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Bufalin alleviates acute kidney injury by regulating NLRP3 inflammasome-mediated pyroptosis

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Abstract

Background

Recently, there has been an increasing clinical incidence of acute kidney injury (AKI), which rapidly declines renal function and leads to massive tubular cell necrosis. Pyroptosis is an inflammatory process of cell death that is more rapid than apoptosis, which is accompanied by a massive release of inflammasome activation. In the study, we aim to explore whether Bufalin regulates the AKI through the pyroptosis pathway.

Methods

We have established gentamicin (GM)-induced AKI animal and cell models to simulate the pathological conditions of kidney injury. The expression of renal injury and pyroptosis-related indicators were detected by western blot. PAS staining and IHC staining were used to analyze renal function. CCK-8 assay was performed to detect cell viability after AKI with different treatments. TUNEL staining, flow cytometry and immunofluorescence assays were performed to measure pyroptosis.

Results

After intraperitoneal injection of GM in rats, renal function was significantly decreased, along with a significant increase of damaged and necrotic cells as suggested by renal tubular epithelial tissue sections. In addition, there was an increase in the pyroptosis-related markers expression and pyroptosis-induced cell death. Consistently, studies in vitro found that GM significantly induced pyroptosis and its associated protein expression in NRK52e cells. Whereas, the administration of Bufalin reversed these effects of GM in vivo and in vitro. Further, we found that Nigericin (NLRP3 agonist) could reversed the effects of bufalin on GM-induced pyroptosis.

Conclusion

Bufalin attenuates pyroptosis generated AKI by inhibiting NLRP3 inflammasome.

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Data Availability

Data sharing not applicable to this article as no datasets were generated or analysed during the current study.

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Funding

Supported by Elderly health Research Project of Jiangsu province 2021(No.LK2021009)and Science and Technology Development Project of Nanjing medical university (NMUB2020049).

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Authors and Affiliations

  1. Department of Pediatrics, The Fourth Affiliated Hospital of Nanjing Medical University, Nanjing, China

    Ling Ding, Xueping Xu, Dan Xie, Dongxiao Xiang & Pingfan Tong

  2. Infectious Department, The Fourth Affiliated Hospital of Nanjing Medical University, Nanjing, China

    Jie Yin

  3. Department of General Practice, The Fourth Affiliated Hospital of Nanjing Medical University, 298 Nanpu road, Jiangbei New District, Nanjing, Jiangsu, China

    Shuyu Liu & Xilan Yang

Authors
  1. Ling Ding
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  2. Jie Yin
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  3. Xueping Xu
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  4. Dan Xie
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  5. Dongxiao Xiang
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  6. Pingfan Tong
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  7. Shuyu Liu
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  8. Xilan Yang
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Contributions

Conceived the hypothesis: L.D. and X.Y.; performed the experiments: L.D., X.X., D.X., D.X., P.T. and S.L.; designed and interpreted the results: L.D., X.X. and X.Y.; wrote the manuscript: L.D. and X.Y.; All authors revised the article critically for important intellectual content. All authors consent for publication.

Corresponding author

Correspondence to Xilan Yang.

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This research was approved by the ethics community of The Fourth Affiliated Hospital of Nanjing Medical University.

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Ding, L., Yin, J., Xu, X. et al. Bufalin alleviates acute kidney injury by regulating NLRP3 inflammasome-mediated pyroptosis. Apoptosis 28, 539–548 (2023). https://doi.org/10.1007/s10495-023-01815-7

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  • DOI: https://doi.org/10.1007/s10495-023-01815-7

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