Double-Negative T Cells Regulate Hepatic Stellate Cell Activation to Promote Liver Fibrosis Progression via NLRP3.

Abstract

Aim:

We mainly explored the role and mechanism of double-negative T cells (DNTs) in liver fibrosis.

Methods:

DNTs were co-cultured with mouse hepatic stellate cells (HSCs). Later, cell viability was detected by Cell Counting Kit-8 (CCK-8) assay; α-SMA expression was measured through fluorescence staining; TNF-α, IL-6, and MMP-9 levels were measured by ELISA; and the expression of Bcl-2, TGF-β1, NLRP3, ASC, and TNFR1 proteins in HSCs was detected by Western blotting (WB) assay. At the same time, HSC-NLRP3-/- and HSC-TNFR1-/- are used to explore the mechanism. In mouse experiments, mice were intraperitoneally injected with DNTs; afterward, the hepatic tissue fibrosis degree was detected by Masson staining, α-SMA expression was measured through immunohistochemistry (IHC) assay, and histopathological changes were detected by sirius-red staining and H&E staining.

Results:

The results suggested that DNTs promoted HSC activation and NLRP3 activation. The effect of DNTs on activating HSC-NLRP3-/- was suppressed, and the difference was significant as compared with HSCs. HSC-TNFR1-/- activation was also inhibited. To explore the mechanism of DNT-secreted TNF-α in TNFR1-NLRP3 activation, we transfected DNTs with TNF-α siRNA; as a result, DNTs with TNF-α silencing did not significantly affect HSC activation. DNTs promoted hepatic tissue fibrosis progression and HSC activation; after treatment with NLRP3 inhibitor, the effect of DNTs on promoting fibrosis was suppressed.

Conclusion:

We discovered that DNTs played an important role in liver fibrosis and that DNTs promoted HSC activation via the TNF-α-TNFR1-NLRP3 signal axis, thus further promoting liver fibrosis progression.

• Authors: Yi Yang, Yongjia Sheng, Jin Wang, Xiaohong Zhou, Wenyan Li, Caiqun Zhang, Li Guo, Chenyang Han
• Journal: Frontiers in immunology (Front Immunol) Vol. 13 Pg. 857116 ( 2022) ISSN: 1664-3224 [Electronic] Switzerland
• PMID: 35371052 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2022 Yang, Sheng, Wang, Zhou, Li, Zhang, Guo and Han.

Chemical References

• NLR Family, Pyrin Domain-Containing 3 Protein
• Nlrp3 protein, mouse
• Receptors, Tumor Necrosis Factor, Type I
• Tumor Necrosis Factor-alpha

Topics

• Animals
• Hepatic Stellate Cells (metabolism)
• Liver Cirrhosis (pathology)
• Mice
• NLR Family, Pyrin Domain-Containing 3 Protein (genetics, metabolism)
• Receptors, Tumor Necrosis Factor, Type I (genetics, metabolism)
• T-Lymphocytes (metabolism)
• Tumor Necrosis Factor-alpha (metabolism)