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Cystine crystal-induced reactive oxygen species associated with NLRP3 inflammasome activation: implications for the pathogenesis of cystine calculi

  • Urology - Original Paper
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Abstract

Purpose

To investigate whether cystine crystal-induced production of reactive oxygen species (ROS) and activation of NLRP3 inflammasome contribute to cystine calculi formation.

Methods

Slc7a9-knockout rats were created as cystine calculi animal models. Kidney histological examination using TEM and immunohistochemistry were performed. The protein expression of NLRP3 and IL-1β and the concentrations of oxidative stress markers such as ROS, MDA and H2O2 in kidney tissues were estimated. In parallel, HK-2 human renal proximal tubule cells were exposed to cystine crystals and NAC treatment. The protein and mRNA expression levels of NLRP3 were evaluated. Finally, cell apoptosis and cystine crystal adherence were also assessed.

Results

Activation of the NLRP3 inflammasome and marked elevations in MDA, H2O2 and ROS levels were observed both in vivo and in vitro. In particular, the protein and mRNA expression of NLRP3 was significantly increased by cystine crystals, but could be restored by an inhibitor of ROS. In addition, cell apoptosis and cystine crystal adherence were promoted by the NLRP3 inflammasome. The expression of CD44, OPN and HA in HK-2 cells was markedly increased by cystine crystals, but could be decreased by NLRP3 siRNA treatment.

Conclusion

Notably, we found that the activation of NLRP3 by cystine crystal-induced ROS production was of major importance in the pathogenesis of cystine calculi formation.

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Data availability

The data used to support the findings of this study are available from the corresponding author upon request.

References

  1. Claes DJ, Jackson E (2012) Cystinuria: mechanisms and management. Pediatr Nephrol 27(11):2031–2038

    Article  PubMed  Google Scholar 

  2. Andreassen KH, Pedersen KV, Osther SS, Jung HU, Lildal SK, Osther PJ (2016) How should patients with cystine stone disease be evaluated and treated in the twenty-first century? Urolithiasis 44(1):65–76

    Article  CAS  PubMed  Google Scholar 

  3. Prencipe G, Caiello I, Cherqui S, Whisenant T, Petrini S, Emma F, De Benedetti F (2014) Inflammasome activation by cystine crystals: implications for the pathogenesis of cystinosis. J Am Soc Nephrol 25(6):1163–1169

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Erickson SB, Vrtiska TJ, Canzanello VJ, Lieske JC (2011) Cystone® for 1 year did not change urine chemistry or decrease stone burden in cystine stone formers. Urol Res 39(3):197–203

    Article  PubMed  Google Scholar 

  5. Mulay SR, Evan A, Anders HJ (2014) Molecular mechanisms of crystal-related kidney inflammation and injury Implications for cholesterol embolism, crystalline nephropathies and kidney stone disease. Nephrol Dial Transplant 29(3):507–514

    Article  CAS  PubMed  Google Scholar 

  6. Knauf F, Asplin JR, Granja I, Schmidt IM, Moeckel GW, David RJ, Flavell RA, Aronson PS (2013) NALP3-mediated inflammation is a principal cause of progressive renal failure in oxalate nephropathy. Kidney Int 84(5):895–901

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Darisipudi MN, Knauf F (2016) An update on the role of the inflammasomes in the pathogenesis of kidney diseases. Pediatr Nephrol 31(4):535–544

    Article  PubMed  Google Scholar 

  8. Komada T, Muruve DA (2019) The role of inflammasomes in kidney disease. Nat Rev Nephrol 15(8):501–520

    Article  PubMed  Google Scholar 

  9. Elliott EI, Sutterwala FS (2015) Initiation and perpetuation of NLRP3 inflammasome activation and assembly. Immunol Rev 265(1):35–52

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Gross O, Thomas CJ, Guarda G, Tschopp J (2011) The inflammasome: an integrated view. Immunol Rev 243(1):136–151

    Article  CAS  PubMed  Google Scholar 

  11. Tsai PY, Ka SM, Chang JM, Lai JH, Dai MS, Jheng HL, Kuo MT, Chen P, Chen A (2012) Antroquinonol differentially modulates T cell activity and reduces interleukin-18 production, but enhances Nrf2 activation, in murine accelerated severe lupus nephritis. Arthritis Rheum 64(1):232–242

    Article  CAS  PubMed  Google Scholar 

  12. Yifan Z, Luwei X, Kai L, Liuhua Z, Yuzheng G, Ruipeng J (2019) Protective effect of salvianolic acid B against oxidative injury associated with cystine stone formation. Urolithiasis 47(6):503–510

    Article  PubMed  Google Scholar 

  13. Liu Y, Duan C, Chen H, Wang C, Liu X, Qiu M, Tang H, Zhang F, Zhou X, Yang J (2018) Inhibition of COX-2/mPGES-1 and 5-LOX in macrophages by leonurine ameliorates monosodium urate crystal-induced inflammation. Toxicol Appl Pharmacol 15(351):1–11

    Google Scholar 

  14. Fernandes MJ, Naccache PH (2018) The role of inhibitory receptors in monosodium urate crystal-induced inflammation. Front Immunol 20(9):1883

    Article  Google Scholar 

  15. Anders HJ, Muruve DA (2011) The inflammasomes in kidney disease. J Am Soc Nephrol 22(6):1007–1018

    Article  CAS  PubMed  Google Scholar 

  16. Martinon F, Pétrilli V, Mayor A, Tardivel A, Tschopp J (2006) Gout-associated uric acid crystals activate the NALP3 inflammasome. Nature 440(7081):237–241

    Article  CAS  PubMed  Google Scholar 

  17. Mulay SR, Kulkarni OP, Rupanagudi KV, Migliorini A, Darisipudi MN, Vilaysane A, Muruve D, Shi Y, Munro F, Liapis H, Anders HJ (2013) Calcium oxalate crystals induce renal inflammation by NLRP3-mediated IL-1β secretion. J Clin Invest 123(1):236–246

    Article  CAS  PubMed  Google Scholar 

  18. Asselman M, Verhulst A, De Broe ME, Verkoelen CF (2003) Calcium oxalate crystal adherence to hyaluronan-, osteopontin-, and CD44-expressing injured/regenerating tubular epithelial cells in rat kidneys. J Am Soc Nephrol 14(12):3155–3166

    Article  CAS  PubMed  Google Scholar 

  19. Verkoelen CF, Van Der Boom BG, Romijn JC (2000) Identification of hyaluronan as a crystal-binding molecule at the surface of migrating and proliferating MDCK cells. Kidney Int 58(3):1045–1054

    Article  CAS  PubMed  Google Scholar 

  20. Vervaet BA, D’Haese PC, De Broe ME, Verhulst A (2009) Crystalluric and tubular epithelial parameters during the onset of intratubular nephrocalcinosis: illustration of the “fixed particle” theory in vivo. Nephrol Dial Transplant 24(12):3659–3668

    Article  CAS  PubMed  Google Scholar 

  21. Verkoelen CF (2006) Crystal retention in renal stone disease: a crucial role for the glycosaminoglycan hyaluronan? J Am Soc Nephrol 17(6):1673–1687

    Article  CAS  PubMed  Google Scholar 

  22. Alelign T, Petros B (2018) Kidney stone disease: an update on current concepts. Adv Urol 2018:3068365

    Article  PubMed  PubMed Central  Google Scholar 

  23. Aggarwal KP, Narula S, Kakkar M, Tandon C (2013) Nephrolithiasis: molecular mechanism of renal stone formation and the critical role played by modulators. Biomed Res Int 2013:292953

    Article  PubMed  PubMed Central  Google Scholar 

  24. Chi T, Taylor E, Stoller ML (2014) Treating the cystine stone former presents a singular clinical challenge. Transl Androl Urol 3(3):234

    PubMed  PubMed Central  Google Scholar 

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Funding

This work was supported by grants from the National Natural Science Foundation of China (NSFC) (Grant Numbers 81570613 and 81370853), Medical research project of Jiangsu Commission of Health (Grant Numbers Z2021074) and Xinghuo Talent Program of Nanjing First Hospital.

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Authors and Affiliations

  1. Department of Urology, Nanjing First Hospital, Nanjing Medical University, 68 Changle Road, Nanjing, 210006, China

    Zhang Yifan, Chen Wei, Xu Luwei, Xu Zheng & Jia Ruipeng

  2. Department of Urology, The Second Affiliated Hospital of Nanjing Medical University, 121#, Jiangjiayuan, Nanjing, 210000, China

    Shen Luming

Authors
  1. Zhang Yifan
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  2. Shen Luming
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  3. Chen Wei
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  4. Xu Luwei
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  5. Xu Zheng
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  6. Jia Ruipeng
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Contributions

ZYF, SLM and CW were responsible for the conception and design, collection and/or assembly of data, manuscript writing, and final approval of the manuscript. XLW was responsible for the data analysis, manuscript writing and final approval of the manuscript. XZ performed the experiments and data collection. JRP was responsible for the conception and design, manuscript correction, financial and administrative support, and final approval of the manuscript. ZY, SL and CW are co-first authors.

Corresponding author

Correspondence to Jia Ruipeng.

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Yifan, Z., Luming, S., Wei, C. et al. Cystine crystal-induced reactive oxygen species associated with NLRP3 inflammasome activation: implications for the pathogenesis of cystine calculi. Int Urol Nephrol 54, 3097–3106 (2022). https://doi.org/10.1007/s11255-022-03347-6

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