中国药物警戒 ›› 2025, Vol. 22 ›› Issue (5): 517-522.
DOI: 10.19803/j.1672-8629.20250036

• 免疫细胞介导的药物性肝损伤研究专栏 • 上一篇    下一篇

雷公藤甲素肝毒性中NK细胞的动态免疫表型研究

陈润吉1, 孙小蕊1, 黄鑫1, 江振洲1, 张陆勇1,2, 王欣之1,*, 丁健3#   

  1. 1中国药科大学多靶标天然药物全国重点实验室新药筛选与药效评价中心,江苏 南京 210009;
    2广东药科大学新药研发中心,广东 广州 510006;
    3南京中医药大学镇江附属医院/镇江市中医院,镇江市中医脾胃病临床医学研究中心,江苏 镇江 212003
  • 收稿日期:2025-01-16 出版日期:2025-05-15 发布日期:2025-05-19
  • 通讯作者: *王欣之,女,副研究员·博导,分子药理学与毒理学。E-mail: wangxz@cpu.edu.cn;#为共同通信作者。
  • 作者简介:陈润吉,女,在读硕士,分子药理学与毒理学。
  • 基金资助:
    国家自然科学基金资助项目（81703626、82073948、82274200、82074114、82174072）; 南京中医药大学镇江附属医院/镇江市中医院,镇江市中医脾胃病临床医学研究中心资助项目(SSPWB2023-KF08,镇江市科技局SS2021005)

Dynamic Immunophenotyping of NK Cells in Triptolide-Induced Hepatotoxicity

CHEN Runji1, SUN Xiaorui1, HUANG Xin1, JIANG Zhenzhou1, ZHANG Luyong1,2, WANG Xinzhi1,*, DING Jian3#   

  1. 1New Drug Screening and Pharmacodynamics Evaluation Center, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing Jiangsu 210009, China;
    2Center for Drug Research and Development, Guangdong Pharmaceutical University, Guangzhou Guangdong 510006, China;
    3Zhenjiang Hospital Affiliated to Nanjing University of Chinese Medicine, Zhenjiang Hospital of Traditional Chinese Medicine, Zhenjiang Traditional Chinese Medicine Spleen and Stomach Disease Clinical Medicine Research Center, Zhenjiang Jiangsu 212003, China
  • Received:2025-01-16 Online:2025-05-15 Published:2025-05-19

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摘要： 目的 探讨NK细胞在雷公藤甲素（TP）肝损伤过程中的动态变化及作用。方法 C57BL/6雌性小鼠分为对照组,TP给药（600 μg·kg-1）1、3、5、7 d组,抗NK1.1抗体与TP合用1、3、5、7 d组。在不同时间点收集小鼠血液,分离肝脏非实质细胞,通过流式细胞术检测NK细胞、CD4+/CD8+T细胞亚群及细胞因子（IFN-γ、IL-4）的时空变化。结果 TP处理后,肝脏NK细胞比例在1 d显著升高（P<0.05）,3~7 d 时CD69+NK细胞持续增加（P<0.05）。细胞因子分泌呈现时间依赖性：5 d时IFN-γ+NK细胞比例升高（P<0.05）,而1和7 d以IL4+NK细胞为主（P<0.05）。NK细胞耗竭显著降低血清丙氨酸氨基转移酶（Alanie Transaminase, ALT）水平（P<0.05）,减少IFN-γ分泌（P<0.05）,使肝脏NK细胞分泌的细胞因子向Th2偏倚（IFN-γ/IL-4比值降低,P<0.05）。此外,TP诱导外周血中CD4+T细胞持续增加（1~7 d均P<0.05）与CD8+T细胞选择性减少（3~5 d, P<0.05）,导致CD4+/CD8+比率失衡。结论 肝脏NK细胞在TP肝损伤中发挥作用。

关键词: 雷公藤甲素, 肝毒性, 自然杀伤细胞, T细胞亚群, 细胞因子, 免疫表型分析, 小鼠

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

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