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Volume 41 Issue 3
Mar.  2025
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Article Navigation >  Journal of Clinical Hepatology  > 2025  >  41(3): 461-468

Characteristics of T cells in the liver tissues of mice with nonalcoholic steatohepatitis

DOI: 10.12449/JCH250311
  • 1.

    Department of Gastroenterology，Shanghai East Hospital，Tongji University，Shanghai 200120，China

  • 2.

    Department of Gastroenterology，Shanghai General Hospital，Shanghai Jiao Tong University，Shanghai 200080，China

Research funding:

Medical Discipline Construction Project of Pudong Health Committee of Shanghai (PWYgf2021-02)

More Information
  • Corresponding author: WANG Jiayi， 18017028830@163.com （ORCID： 0009-0004-8097-5288）
  • Received Date: 2024-07-24
  • Accepted Date: 2024-09-23
  • Published Date: 2025-03-25
    Abstract
  •   Objective  To investigate the heterogeneity and transcriptomic characteristics of T-cell subsets in the liver of mice with nonalcoholic steatohepatitis （NASH） at the single-cell level using single-cell RNA sequencing （scRNA-seq）， and to provide a reference for studying the mechanism of action of T cells in NASH.  Methods  Six male C57BL/6 mice were randomly divided into control group fed with regular diet and NASH group fed with methionine-choline-deficient （MCD） diet， with three mice in each group， and liver tissue was collected for scRNA-seq after 6 weeks of modeling. Specific differentially expressed genes were analyzed between T-cell subsets， and related analyses were performed， including dimension clustering， cell type annotation， t-distributed stochastic neighbor embedding （t-SNE）， violin plot， gene ontology （GO） functional enrichment analysis， and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis. Immunofluorescent staining was used to observe the expression of the T cell marker Tcrα and the specific marker genes Tcf7 and Cxcr6 in the liver of mice in the two groups. The independent-samples t test was used for comparison of continuous data between two groups.  Results  Two T cell subsets were identified in the liver of mice， and the percentage of cluster 6 decreased from 58.5% in the control group to 48.7% in the NASH group. The top four specific genes were Nsg2， Cd8b1， Cd8a， and Tcf7. Tcf7， a characteristic marker gene for cluster 6， was expressed in 65% of cells in cluster 6， and therefore， cluster 6 was defined as Tcf7+ T cells. The GO and KEGG enrichment analyses showed that the differentially expressed genes of cluster 6 were involved in T cell activation， leukocyte adhesion， binding ubiquitin-like protein ligase， and the signaling pathways for Th17， Th1， and Th2 cell differentiation. The percentage of cluster 7 increased from 41.5% in the control group to 51.3% in the NASH group. The top four specific genes of cluster 7 were Cd40lg， Tcrg-C1， Il2rα， and Cxcr6. Cxcr6 was expressed in 90% of cells in cluster 7， and therefore， cluster 7 was defined as Cxcr6+ T cells. The GO and KEGG enrichment analyses showed that cluster 7 was involved in T cell activation， cytokine production， the T cell receptor signaling pathway， and the Th17 cell differentiation and MAPK signaling pathway. Immunofluorescence assay showed that compared with the control group， the NASH group showed a significant reduction in the area with positive co-expression of Tcf7 protein and Tcrα protein （1.80%±0.67% vs 0.33%±0.13%， P<0.05） and a significant increase in the area with positive co-expression of Cxcr6 protein and Tcrα protein （0.50%±0.09% vs 2.66%± 0.33%， P<0.001）.  Conclusion  There is a reduction in the percentage of Tcf7+ T cells and an increase in the percentage of Cxcr6+ T cells in NASH mice， revealing the characteristics and differences of T cells in the liver of NASH mice.

     

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