Skip to main content
SpringerLink
Log in

Plasma exosomes derived from patients with intestinal Behçet’s syndrome induce intestinal epithelial cell pyroptosis

  • Original Article
  • Published:
Clinical Rheumatology Aims and scope Submit manuscript

Abstract

Objectives

Intestinal Behçet’s syndrome (IBS) has high morbidity and mortality rates with serious complications. The purpose of this study was to investigate the expression of pyroptosis-related proteins in the intestinal tissues of IBS patients and explore the role of plasma exosomes derived from IBS patients in the pyroptosis of intestinal epithelial cells.

Method

Immunohistochemistry was used to investigate the expression of nucleotide-binding domain-like receptor protein 3 (NLRP3), caspase-1, and gasdermin D (GSDMD). Quantitative real-time PCR was employed to measure the mRNA levels of IL-1β and IL-18 in the intestinal tissues. Plasma exosomes were isolated and observed by transmission electron microscopy. The exosomes were co-cultured with intestinal epithelial cells in vitro. Western blot was used to measure the expression of pyroptosis-related proteins including NLRP3, full-length GSDMD, N-terminal GSDMD, pro-caspase-1, and cleaved caspase-1. The levels of IL-1β and IL-18 were detected by enzyme-linked immunosorbent assay. Cell death was measured by using the lactate dehydrogenase (LDH) release assay.

Results

Expression of NLRP3 (12.2% ± 1.2%, 8.1% ± 0.9%, t = 4.692, p = 0.009), caspase-1 (24.6% ± 2.1%, 4.2% ± 1.8%, t = 12.842, p = 0.000), and GSDMD (16.6% ± 1.9%, 9.8% ± 1.3%, t = 5.194, p = 0.007) were significantly increased in the intestinal tissues of patients with IBS compared with normal control (NC) group, respectively. The relative mRNA levels of IL-1β (t = 4.308, p = 0.005) and IL-18 (t = 3.096, p = 0.021) in the intestinal tissues were significantly higher in IBS patients than in NC group, while the protein levels of IL-1β (t = 3.873, p = 0.018) and IL-18 (t = 4.389, p = 0.012) were also significantly increased, which was consistent with the results of the relative mRNA levels. Moreover, we found that exosomes from IBS patients significantly induced pyroptosis of intestinal epithelial cells via the activation of NLRP3 inflammasome in vitro experiments.

Conclusions

Plasma exosomes derived from IBS patients may induce pyroptosis of intestinal epithelial cells via the activation of NLRP3 inflammasome.

Key Points

•The role of exosomes in IBS is first reported in this study.

• In this study, we explored the mechanism that plasma exosomes derived from IBS patients may induce pyroptosis of intestinal epithelial cells via the activation of NLRP3 inflammasome.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Data availability

Yes.

Code availability

Not applicable.

References

  1. Sakane T, Takeno M, Suzuki N, Inaba G (1999) Behcet’s disease. N Engl J Med 341:1284–1291

    Article  CAS  Google Scholar 

  2. Hatemi I, Hatemi G, Celik AF (2018) Gastrointestinal involvement in Behcet disease. Rheum Dis Clin N Am 44:45–64

    Article  Google Scholar 

  3. Zou J, Luo JF, Shen Y, Cai JF, Guan JL (2021) Cluster analysis of phenotypes of patients with Behçet's syndrome: a large cohort study from a referral center in China. Arthritis Res Ther 23(1):45

    Article  Google Scholar 

  4. Smith JA, Siddiqui D (2002) Intestinal Behçet’s disease presenting as a massive acute lower gastrointestinal bleed. Dig Dis Sci 47:517–521

    Article  Google Scholar 

  5. Lu F, Lan Z, Xin Z, He C, Guo Z, Xia X, Hu T (2020) Emerging insights into molecular mechanisms underlying pyroptosis and functions of inflammasomes in diseases. J Cell Physiol 235(4):3207–3021

    Article  CAS  Google Scholar 

  6. Shi J, Gao W, Shao F (2017) Pyroptosis: gasdermin-mediated programmed necrotic cell death. Trends Biochem 42:245–254

    Article  CAS  Google Scholar 

  7. Isabella R, Katherine AD, Daisy XJ, von Jakob M, Jeannette LT, Angus YL, Naomi HP, Janelle SA, Igor EB, Karsten G, Russell EV (2017) NAIP-NLRC4 inflammasomes coordinate intestinal epithelial cell expulsion with eicosanoid and IL-18 release via activation of caspase-1 and -8. Immunity 46:649–659

    Article  Google Scholar 

  8. Li YY, Xia WW, Wu MY, Yin J, Wang Q, Li SZ, Zhang AH, Huang SM, Zhang Y, Jia ZJ (2020) Activation of GSDMD contributes to acute kidney injury induced by cisplatin. Am J Physiol Ren Physiol 318:F96–F106

    Article  CAS  Google Scholar 

  9. Li Z, Guo J, Bi L (2020) Role of the NLRP3 inflammasome in autoimmune diseases. Biomed Pharmacother 130:110542

    Article  CAS  Google Scholar 

  10. Swanson KV, Deng M, Ting JP (2019) The NLRP3 inflammasome: molecular activation and regulation to therapeutics. Nat Rev Immunol 19:477–489

    Article  CAS  Google Scholar 

  11. Shen HH, Yang YX, Meng X, Luo XY, Li XM, Shuai ZW, Ye DQ, Pan HF (2018) NLRP3: a promising therapeutic target for autoimmune diseases. Autoimmun Rev 17:694–702

    Article  CAS  Google Scholar 

  12. Kim EH, Park MJ, Park S, Lee ES (2015) Increased expression of the NLRP3 inflammasome components in patients with Behçet’s disease. J Inflamm (Lond) 12:41

    Article  Google Scholar 

  13. Kalluri R, LeBleu VS (2019) Exosomes. Annu Rev Biochem 88:487–514

    Article  Google Scholar 

  14. Zhu T, Wang Y, Jin H, Li L (2019) The role of exosome in autoimmune connective tissue disease. Ann Med 51(2):101–108

    Article  CAS  Google Scholar 

  15. Watanabe K, Tanida S, Inoue N, Kunisaki R, Kobayashi K, Nagahori M, Arai K, Uchino M, Koganei K, Kobayashi T, Takeno M, Ueno F, Matsumoto T, Mizuki N, Suzuki Y, Hisamatsu T (2020) Evidence-based diagnosis and clinical practice guidelines for intestinal Behçet’s disease 2020 edited by Intractable Diseases, the Health and Labour Sciences Research Grants. J Gastroenterol 55(7):679–700

    Article  Google Scholar 

  16. Lawton G, Bhakta BB, Chamberlain MA, Tennant A (2004) The Behcet’s disease activity index. Rheumatology (Oxford) 43:73–78

    Article  CAS  Google Scholar 

  17. Leccese P, Alpsoy E (2019) Behcet’s disease: an overview of etiopathogenesis. Front Immunol 10:1067

    Article  CAS  Google Scholar 

  18. Turner JR (2009) Intestinal mucosal barrier function in health and disease. Nat Rev Immunol 9:799–809

    Article  CAS  Google Scholar 

  19. Bulek K, Zhao JJ, Liao Y, Rana N, Corridoni D, Antanaviciute A, Chen X, Wang H, Qian W, Miller-Little WA, Swaidani S, Tang FQ, Willard BB, McCrae K, Kang ZZ, Dubyak GR, Cominelli F, Simmons A, Pizarro TT, Li XX (2020) Epithelial-derived gasdermin D mediates nonlytic IL-1beta release during experimental colitis. J Clin Invest 130:4218–4234

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Lu F, Lan Z, Xin Z, He C, Guo Z, Xia X, Hu T (2020) Emerging insights into molecular mechanisms underlying pyroptosis and functions of inflammasomes in diseases. J Cell Physiol 235:3207–3221

    Article  CAS  Google Scholar 

  21. Kelley N, Jeltema D, Duan Y, He Y (2019) The NLRP3 inflammasome: an overview of mechanisms of activation and regulation. Int J Mol Sci 20(13):3328

    Article  CAS  Google Scholar 

  22. Kirino Y, Zhou Q, Ishigatsubo Y, Mizuki N, Tugal-Tutkun I, Seyahi E, Özyazgan Y, Ugurlu S, Erer B, Abaci N, Ustek D, Meguro A, Ueda A, Takeno M, Inoko H, Ombrello MJ, Satorius CL, Maskeri B, Mullikin JC, Sun HW, Gutierrez-Cruz G, Kim Y, Wilson AF, Kastner DL, Gül A, Remmers EF (2013) Targeted resequencing implicates the familial Mediterranean fever gene MEFV and the toll-like receptor 4 gene TLR4 in Behçet disease. Proc Natl Acad Sci U S A 110(20):8134–8139

    Article  CAS  Google Scholar 

  23. Burillo-Sanz S, Montes-Cano MA, García-Lozano JR, Olivas-Martínez I, Ortego-Centeno N, García-Hernández FJ, Espinosa G, Graña-Gil G, Sánchez-Bursón J, Juliá MR, Solans R, Blanco R, Barnosi-Marín AC, Gómez de la Torre R, Fanlo P, Rodríguez-Carballeira M, Rodríguez-Rodríguez L, Camps T, Castañeda S, Alegre-Sancho JJ, Martín J, González-Escribano MF (2019) Behçet’s disease and genetic interactions between HLA-B*51 and variants in genes of autoinflammatory syndromes. Sci Rep 9(1):2777

    Article  Google Scholar 

  24. Li L, Yu H, Jiang Y, Deng B, Bai L, Kijlstra A, Yang P (2016) Genetic variations of NLR family genes in Behcet’s disease. Sci Rep 6:20098

    Article  CAS  Google Scholar 

  25. Yüksel Ş, Eren E, Hatemi G, Sahillioğlu AC, Gültekin Y, Demiröz D, Akdiş C, Fresko İ, Özören N (2014) Novel NLRP3/cryopyrin mutations and pro-inflammatory cytokine profiles in Behçet’s syndrome patients. Int Immunol 26(2):71–81

    Article  Google Scholar 

  26. Padula MC, Leccese P, Lascaro N, Padula AA, Carbone T, Martelli G, D'Angelo S (2019) Identification of a de novo NLRP3 gene variation in an Italian Behçet syndrome patient. Int J Immunogenet 46(5):339–341

    Article  CAS  Google Scholar 

  27. Liu Y, Lou G, Li A, Zhang T, Qi J, Ye D, Zheng M, Chen Z (2018) AMSC-derived exosomes alleviate lipopolysaccharide/d-galactosamine-induced acute liver failure by miR-17-mediated reduction of TXNIP/NLRP3 inflammasome activation in macrophages. EBioMedicine 36:140–150

    Article  Google Scholar 

  28. Dai Y, Wang S, Chang S, Ren D, Shali S, Li C, Yang H, Huang Z, Ge J (2020) M2 macrophage-derived exosomes carry microRNA-148a to alleviate myocardial ischemia/reperfusion injury via inhibiting TXNIP and the TLR4/NF-kappaB/NLRP3 inflammasome signaling pathway. J Mol Cell Cardiol 142:65–79

    Article  CAS  Google Scholar 

  29. Kahraman T, Gucluler G, Simsek I, Yagci FC, Yildirim M, Ozen C, Dinc A, Gursel M, Ikromzoda L, Sutlu T, Gay S, Gursel I (2017) Circulating LL37 targets plasma extracellular vesicles to immune cells and intensifies Behcet’s disease severity. J Extracell Vesicles 6:1284449

    Article  Google Scholar 

Download references

Funding

This project was supported by the Clinical Science Innovation Program of Shanghai Shenkang Hospital Development Center (grant No. SHDC12017129) and the General Program of National Natural Science Foundation of China (grant No. 81871276).

Author information

Authors and Affiliations

  1. Department of Rheumatology and Immunology, Huadong Hospital Affiliated to Fudan University, Shanghai, China

    Cheng-cheng Hou, Hai-fen Ma, Jing-fen Ye, Dan Luo, Hua-fang Bao & Jian-long Guan

Authors
  1. Cheng-cheng Hou
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hai-fen Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jing-fen Ye
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Dan Luo
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hua-fang Bao
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jian-long Guan
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Study design and supervision were done by Jian-long Guan; study operation and composition writing were done by Cheng-cheng Hou; blood sample collection was done by Hai-fen Ma and Jing-fen Ye; and data collection and statistical analysis were done by Dan Luo and Hua-fang Bao.

Corresponding author

Correspondence to Jian-long Guan.

Ethics declarations

Ethics approval

The study was approved by the Research Ethics Committee of Huadong Hospital Affiliated to Fudan University (Grant No. 2020K034), and informed consent was signed by all participants.

Consent to participate

Yes.

Consent for publication

Yes.

Disclosures

None.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hou, Cc., Ma, Hf., Ye, Jf. et al. Plasma exosomes derived from patients with intestinal Behçet’s syndrome induce intestinal epithelial cell pyroptosis. Clin Rheumatol 40, 4143–4155 (2021). https://doi.org/10.1007/s10067-021-05755-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10067-021-05755-y

Keywords

Search

Navigation