Dynamic Immunophenotyping of NK Cells in Triptolide-Induced Hepatotoxicity

doi:10.19803/j.1672-8629.20250036

Abstract

Abstract: Objective To explore the dynamic changes and roles of natural killer (NK) cells in the process of triptolide (TP)-induced liver injury. Methods Female C57BL/6 mice were divided into the control group, the 1, 3, 5, and 7 days of TP administration (600 μg·kg^-1) groups, and groups treated with anti-NK1.1 antibody combined with TP for 1, 3, 5, and 7 days. At different time points, the blood of mice was collected and non-parenchymal cells in the liver were isolated for flow cytometry analysis of the temporal changes in NK cells, CD4⁺/CD8⁺ T cell subsets and cytokines (IFN-γ and IL-4). Results After TP treatment, the proportion of liver NK cells significantly increased at day 1 (P<0.05), while CD69⁺ NK cells kept increasing from day 3 to 7(P<0.05). NK cell depletion significantly reduced serum ALT levels (P<0.05) and inhibited IFN-γ production (P<0.05), shifting NK cell-derived cytokines toward a Th2 bias (reduced IFN-γ/IL-4 ratio, P<0.05). Additionally, TP induced a persistent increase in peripheral blood CD4⁺ T cells (days 1-7, P<0.05) and a selective decrease in CD8⁺ T cells (days 3-5, P<0.05), leading to an imbalanced CD4⁺/CD8⁺ ratio. Conclusion Hepatic NK cells can contribute to TP-induced liver injury.

Key words: Triptolide, Hepatotoxicity, Natural Killer Cells, T Cell Subsets, Cytokines, Immunophenotyping, Mice

CLC Number:

R979.5

CHEN Runji, SUN Xiaorui, HUANG Xin, JIANG Zhenzhou, ZHANG Luyong, WANG Xinzhi, DING Jian. Dynamic Immunophenotyping of NK Cells in Triptolide-Induced Hepatotoxicity[J]. Chinese Journal of Pharmacovigilance, 2025, 22(5): 517-522.

Add to citation manager EndNote|Ris|BibTeX

URL: https://zgywjj.magtechjournal.com/EN/10.19803/j.1672-8629.20250036

https://zgywjj.magtechjournal.com/EN/Y2025/V22/I5/517

References

[1] YUAN K, LI X, LU Q, et al.Application and Mechanisms of Triptolide in the Treatment of Inflammatory Diseases-a Review[J]. Front in Pharmacology, 2019, 10: 1469
[2] TANG QH, ZHANG HR, ZHANG LY, et al.Research Progress in Triptolide-Induced Live Injury[J]. Chinese Journal of Pharmαcovigilance(中国药物警戒), 2025, 22(2): 121-127.
[3] ZOU M, NONG C, YU Z, et al.The Role of Invariant Natural Killer T Cells and Associated Immunoregulatory Factors in Triptolide-Induced Cholestatic Liver Injury[J]. Food and Chemical Toxicology, 2020, 146, 111777.
[4] WANG X, XUE R, ZHANG S, et al.Activation of Natural Killer T Cells Contributes to Triptolide-Induced Liver Injury in Mice[J]. Acta Pharmacologica Sinica, 2018, 39(12): 1847-1854.
[5] PING DB, QI J, SUN XS, et al.Protective Effect of All-Trans Retinoic Acid on Natural Killer Cells Damaged by Oxidative Stress in Liver Fibrosis[J]. Chinese Journal of Immunology(中国免疫学杂志), 2024, 40(7): 1359-1363.
[6] QIAN Y, ZHAO J, WU H, et al.Innate Immune Regulation in Inflammation Resolution and Liver Regeneration in Drug-Induced Liver Injury[J]. Archives of Toxicology, 2025, 99(1): 115-126.
[7] ZHANG H, YUAN Z, WANG J, et al.Triptolide Leads to Hepatic Intolerance to Exogenous Lipopolysaccharide and Natural-Killer-Cell Mediated Hepatocellular Damage by Inhibiting MHC Class I Molecules[J]. Phytomedicine. 2023, 109: 154621.
[8] KONG W, LI X, ZOU M, et al.iNKT17 Cells Play a Pathogenic Role in Ethinylestradiol-Induced Cholestatic Hepatotoxicity[J]. Archives of Toxicology, 2023, 97(2): 561-580.
[9] FRIEDRICH C, TAGGENBROCK RLRE, DOUCET-LADEVÈZE R, et al. Effector Differentiation Downstream of Lineage Commitment in ILC1s Is Driven by Hobit across Tissues[J]. Nature Immunology, 2021, 22(10): 1256-1267.
[10] GU M, ZHANG Y, LIN Z, et al.Decrease in UCP1 by Sustained High Lipid Promotes NK Cell Necroptosis to Exacerbate Nonalcoholic Liver Fibrosis[J]. Cell Death & Disease, 2024, 15(7): 518.
[11] CROUSE J, XU HC, LANG PA, et al.NK Cells Regulating T Cell Responses: Mechanisms and Outcome[J]. Trends in Immunology, 2015, 36(1): 49-58.
[12] WANG J, SUN R, WEI H, et al.Pre-Activation of T Lymphocytes by Low Dose of Concanavalin A Aggravates Toll-Like Receptor-3 Ligand-Induced NK Cell-Mediated Liver Injury[J]. International Immunopharmacology, 2006, 6(5): 800-807.
[13] JAESCHKE H, RAMACHANDRAN A.Mechanisms and Pathoph-ysiological Significance of Sterile Inflammation during Acetaminophen Hepatotoxicity[J]. Food and Chemical Toxicology, 2020, 138: 111240.
[14] KINIWA T, ENOMOTO Y, TERAZAWA N, et al.NK Cells Activated by Interleukin-4 in Cooperation with Interleukin-15 Exhibit Distinctive Characteristics[J]. Proceedings of the National Academy of Sciences of the United States of America, 2016, 113(36): 10139-10144.
[15] CHENG C, ZHANG Q, LI Y, et al.Interplay between Liver Type 1 Innate Lymphoid Cells and NK Cells Drives the Development of Alcoholic Steatohepatitis[J]. Cell and Molecular Gastroenterology Hepatology, 2023, 15(1): 261-274.
[16] ZHOU Z, YAO J, WU D, et al.Type 2 Cytokine Signaling in Macro-phages Protects from Cellular Senescence and Organismal Aging[J]. Immunity, 2024, 57(3): 513-527.
[17] DUBROT J, DU PP, LANE-RETICKER SK, et al.In vivo CRISPR Screens Reveal the Landscape of Immune Evasion Pathways across Cancer[J]. Nature Immunology, 2022, 23(10): 1495-1506.
[18] CHEN X, GHANIZADA M, MALLAJOSYULA V, et al.Differential Roles of Human CD4+ and CD8+ Regulatory T Cells in Controlling Self-Reactive Immune Responses[J]. Nature Immunology, 2025, 26(2): 230-239.