临床肝胆病杂志2025，Vol.41Issue(3)：461-468,8.DOI:10.12449/JCH250311
非酒精性脂肪性肝炎小鼠模型肝组织T淋巴细胞的特征分析
Characteristics of T cells in the liver tissues of mice with nonalcoholic steatohepatitis
摘要
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.关键词
非酒精性脂肪性肝炎/T淋巴细胞/单细胞测序/小鼠,近交C57BLKey words
Non-Alcoholic Steatohepatitis/T-Lymphocytes/Single-cell RNA Sequencing/Mice,Inbred C57BL引用本文复制引用
冒婷,徐铭益,王佳轶..非酒精性脂肪性肝炎小鼠模型肝组织T淋巴细胞的特征分析[J].临床肝胆病杂志,2025,41(3):461-468,8.基金项目
上海市浦东新区卫生健康委员会医学学科建设项目(PWYgf2021-02) Medical Discipline Construction Project of Pudong Health Committee of Shanghai(PWYgf2021-02) （PWYgf2021-02）
