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Bone marrow mesenchymal stem cells-derived exosomes containing miR-539-5p inhibit pyroptosis through NLRP3/caspase-1 signalling to alleviate inflammatory bowel disease

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Abstract

Background

Exosomes derived from bone mesenchymal stem cells (BMSCs) are potential candidates for inflammatory bowel disease (IBD) treatment. The present study investigated the therapeutic effect and potential mechanism of BMSCs-derived exosomes on pyroptosis in IBD.

Methods

We induced IBD in mice and cell models through dextran sulfate sodium (DSS) and LPS, respectively. The mRNA and protein expression levels were assessed by qRT–PCR, Western blotting, IF and IHC. The concentrations of IL-1β, IL-18 and TNFα were assessed using ELISA. ROS levels were determined using DCFH-DA staining. Cell proliferation of mIECs was analysed using an MTT assay. In addition, a flow cytometry assay was performed to detect pyroptosis. Finally, the binding relationship between miR-539-5p and NLRP3 was verified by a dual luciferase reporter gene assay.

Results

Our results revealed that intraperitoneal injection of BMSCs-derived exosomes inhibited DSS-induced pyroptosis as well as IBD symptoms in mice. In addition, BMSCs-derived exosome treatment suppressed pyroptosis, ROS levels and the concentrations of proinflammatory cytokines (IL-1β, IL-18 and TNFα) in LPS-treated mIECs in a miR-539-5p-dependent manner. Further research found that miR-539-5p suppressed NLRP3 expression in mIECs by directly targeting NLRP3. As expected, pyroptosis in LPS-treated mIECs was significantly reduced by NLRP3 knockdown. In addition, NLRP3 silencing restored the inhibitory effect of exosomes derived from BMSCs transfected with miR-539-5p inhibitor on pyroptosis in LPS-treated mIECs.

Conclusion

The present study demonstrated that BMSCs-derived exosomal miR-539-5p suppresses pyroptosis through NLRP3/caspase-1 signalling to inhibit IBD progression.

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Abbreviations

IBD:

Inflammatory bowel disease

UC:

Ulcerative colitis

CD:

Crohn’s disease

MCSs:

Mesenchymal stem cells

BMSCs:

Bone marrow mesenchymal stem cells

miRNAs:

MicroRNAs

PYD:

Pyrin domain

NLRP3:

Nucleotide-binding domain-like receptor family member pyrin domain-containing protein 3

ASC:

Apoptosis-associated speck-like protein containing CARD

GSDMD:

Gasdermin D

ROS:

Reactive oxygen species

TEM:

Transmission electron microscopy

MTT:

3-(4,5-Dimethylthiazolyl2)-2,5-diphenyltetrazolium bromide

qRT–PCR:

Quantitative real-time polymerase chain reaction

DSS:

Dextran sulfate sodium salt

TNF-α:

Tumour necrosis factor-α

IL:

Interleukin

LPS:

Lipopolysaccharide

IF:

Immunofluorescence

HE:

Haematoxylin–eosin

SD:

Standard deviation

ANOVA:

Analysis of variance

DMELM:

Dulbecco's modified Eagle’s medium

ELISA:

Enzyme-linked immunosorbent assay

IHC:

Immunohistochemistry

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Acknowledgements

We would like to give our sincere gratitude to the reviewers for their constructive comments.

Funding

This work was supported by Shandong Traditional Chinese Medicine Science and Technology Development Project (No. 2019-0397), Special funding sponsorship of Shandong Province Traditional Chinese Medicine High-level Talent Cultivation Project and Qilu Health Leading Talents Training Project Special Fund.

Author information

Author notes

  1. Dandan Wang and Hui Xue contributed equally to this work.

Authors and Affiliations

  1. Department of Anorectum, The Affiliated Hospital of Qingdao University, No. 16 Jiangsu Road, Qingdao, 266000, Shandong Province, People’s Republic of China

    Dandan Wang, Penglin Liu, Cuixia Qiao, Chengjian Pang & Lize Zhang

  2. Department of Gynecology, Qingdao Hospital of Traditional Chinese Medicine, Qingdao, 266100, Shandong Province, People’s Republic of China

    Hui Xue

  3. Department of Pain Clinic, Qingdao 8th People’s Hospital, Qingdao, 266100, Shandong Province, People’s Republic of China

    Jianfeng Tan

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  1. Dandan Wang
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Correspondence to Lize Zhang.

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Conflicts of interest

All authors agree with the presented findings, have contributed to the work, and declare no conflict of interest.

Ethics approval and consent to participate

The third-generation BMSCs were used for experiments. This study was approved by the The Affiliated Hospital of Qingdao University ethics committee. All animal experiments and protocols were reviewed and approved by the Animal Care and Use Committee of The Affiliated Hospital of Qingdao University.

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Responsible Editor: H. Wang.

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Wang, D., Xue, H., Tan, J. et al. Bone marrow mesenchymal stem cells-derived exosomes containing miR-539-5p inhibit pyroptosis through NLRP3/caspase-1 signalling to alleviate inflammatory bowel disease. Inflamm. Res. 71, 833–846 (2022). https://doi.org/10.1007/s00011-022-01577-z

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  • DOI: https://doi.org/10.1007/s00011-022-01577-z

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