五灵胶囊对刀豆蛋白A诱导自身免疫性肝炎小鼠的保护作用
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国家现代农业产业技术体系资助项目(CARS-21);中药大品种品牌价值提升示范研究项目(202190025);陕西省科技厅重点研发计划项目(2024SF-YBXM-458);陕西省教育厅重点科学研究计划项目(22JY019);咸阳市2020重大科技专项项目(2020K01-20),陕西高校青年创新团队项目(2023).


Protective effects of Wuling capsule on concanavalin A-induced autoimmune hepatitis mice
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Supported by Project of National Modern Agricultural Industrial Technology System (CARS-21);Demonstration Research Project on Brand Value Enhancement of Large Chinese Medicine Varieties (202190025);Key Research and Development Project of Science and Technology Department of Shaanxi Province (2024SF-YBXM-458);Key Project of Scientific Research of Education Department of Shaanxi (22JY019);2020 Major Science and Technology Project of Xianyang (2020K01-20), and Shaanxi University Youth Innovation Team Project (2023).

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    摘要:

    目的 探讨五灵胶囊对自身免疫性肝炎(AIH)小鼠的保护作用。方法 小鼠随机分为对照组、AIH模型组、五灵胶囊低剂量组(0.5 g·kg-1·d-1)、五灵胶囊中剂量组(1.0 g·kg-1·d-1)、五灵胶囊高剂量组(2.0 g·kg-1·d-1),每组10只。五灵胶囊各剂量组按10 mL/kg的体积灌胃给予五灵胶囊混悬液,每天1次;对照组和AIH模型组灌胃给予等体积生理盐水。给药14 d后,AIH模型组和五灵胶囊各剂量组小鼠尾静脉注射刀豆蛋白A(20 mg/kg),注射8 h后收集血清、肝脏和脾脏。采用全自动生化分析仪检测血清丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)水平,用H-E染色观察肝组织病理结构,用ELISA法测定肝组织中IL-6、IL-4和TNF-α含量,用qPCR法测定肝组织中IL-6IL-4TNF-α、Toll样受体4(TLR4)和NF-κB mRNA的相对表达量,用免疫荧光法分析肝组织中NF-κB p65蛋白表达和核移位情况。结果 与对照组相比,AIH模型组小鼠肝脏形态结构异常,血清ALT和AST水平升高,肝组织中IL-4、IL-6和TNF-α含量升高,肝组织中IL-6IL-4TNF-αTLR4NF-κB mRNA的相对表达量上调,NF-κB p65的入核增加。五灵胶囊明显改善AIH模型小鼠的肝脏病理结构,降低血清ALT和AST水平,降低肝组织中IL-4、IL-6和TNF-α含量及TLR4NF-κBIL-4IL-6 mRNA的相对表达量,并抑制NF-κB p65的入核激活。结论 五灵胶囊对AIH小鼠具有明显的保护作用,其机制可能与TLR4/NF-κB信号通路有关。

    Abstract:

    Objective To investigate the protective effects of Wuling capsule on mice with autoimmune hepatitis (AIH). Methods Mice were randomly divided into control group, AIH model group, Wuling capsule low-dose group (0.5 g·kg-1·d-1), Wuling capsule middle-dose group (1.0 g·kg-1·d-1), and Wuling capsule high-dose group (2.0 g·kg-1·d-1), with 10 mice in each group. The Wuling capsule groups were administered with Wuling capsule suspension of different doses orally at a volume of 10 mL/kg once daily; the control group and AIH model group were given the same volume of saline by gavage. After 14 d of administration, mice in the AIH model group and Wuling capsule groups were injected with concanavalin A (20 mg/kg) via the tail vein, and the serum, liver, and spleen were collected 8 h after injection. Serum alanine transaminase (ALT) and aspartate transaminase (AST) levels were measured using an automatic biochemical analyzer; hematoxylin-eosin staining was used to observe the pathological structure of the liver tissue; the contents of interleukin (IL)-6, IL-4, and tumor necrosis factor (TNF)-α in liver were determined by enzyme-linked immunosorbent assay, and quantitative polymerase chain reaction was used to determine the relative mRNA expression levels of IL-6, IL-4, TNF-α, Toll-like receptor 4 (TLR4), and nuclear factor κB (NF-κB) in the liver. Fluorescence immunoassay was used to analyze the expression and nuclear translocation of NF-κB p65 protein in the liver. Results Compared with the control group, the AIH model group showed abnormal liver morphology and structure, increased serum ALT and AST levels, increased contents of IL-4, IL-6, and TNF-α in the liver, upregulated mRNA expression levels of IL-6, IL-4, TNF-α, TLR4, and NF-κB in the liver, and increased nuclear entry of NF-κB p65. Wuling capsule significantly improved the pathological structure of the liver in AIH mice, reduced serum ALT and AST levels, decreased the contents of IL-4, IL-6, and TNF-α in the liver and the mRNA expression levels of TLR4, NF-κB, IL-4, IL-6, and inhibited the nuclear activation of NF-κB p65. Conclusion Wuling capsule has significant protective effects on AIH mice, which may be related to the TLR4/NF-κB signaling pathway.

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  • 收稿日期:2023-02-17
  • 最后修改日期:2024-02-20
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  • 在线发布日期: 2025-01-17
  • 出版日期: 2025-01-20
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