Value of Fuzheng Huayu prescription in preventing liver fibrosis by altering the phenotypic function of CD8<sup>+</sup> T lymphocytes in the liver of mice with acute liver injury

Hui HUANG; Lieming XU; Jian PING; Yang ZHOU

doi:10.3969/j.issn.1001-5256.2022.02.017

Volume 38 Issue 2

Feb. 2022

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Article Navigation > Journal of Clinical Hepatology > 2022 > 38(2): 342-346

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Value of Fuzheng Huayu prescription in preventing liver fibrosis by altering the phenotypic function of CD8⁺ T lymphocytes in the liver of mice with acute liver injury

DOI: 10.3969/j.issn.1001-5256.2022.02.017

Hui HUANG^{1, 2},
Lieming XU^{1, 2, 3},
Jian PING^{1, 4},
Yang ZHOU^{5
,
,}

1.
Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
2.
Institute of Liver Diseases, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
3.
Key Laboratory of Liver and Kidney Diseases, Ministry of Education, Shanghai 201203, China
4.
Institute of Brain Science, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
5.
Preventive Treatment center Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200437, China

Research funding:

National Natural Science Foundation of China (81573794)

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Corresponding author: ZHOU Yang, 973kt@sina.com
Received Date: 2021-06-26
Accepted Date: 2021-08-12
Published Date: 2022-02-20

Abstract

Abstract

Objective To investigate the effect of liver CD8⁺ T lymphocytes on co-cultured hepatic stellate cells (HSCs) after the application of Fuzheng Huayu prescription in a moues model of acute liver injury, as well as the mechanism of action of Fuzheng Huayu prescription in preventing liver fibrosis. Methods A total of 18 specific pathogen-free male C57BL/6NCrl Vr mice were randomly divided into normal group, model group, and Fuzheng Huayu prescription group, with 6 mice in each group. The mice in the Fuzheng Huayu prescription group were given Fuzheng Huayu prescription for 5 days in advance. At 12 hours before the experiment, 10% CCl₄ was injected intraperitoneally at a dose of 2 mL/kg body weight. Blood was collected from the main abdominal vein, and the serum was separated to measure the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Part of the liver was used for pathological observation. After the mice were pretreated with medication in vivo, flow cytometry was used for the sorting of mouse liver CD8⁺ T lymphocytes, which were then co-cultured with the mouse HSC cell line (JS 1) in a 96-well plate at a ratio of 2∶ 1, and after co-culture for 24 and 48 hours, qPCR was used to measure the changes in the mRNA expression of Col.I and α-SMA. An analysis of variance was used for comparison of normally distributed continuous data between multiple groups, and the SNK-q test or the least significant difference t-test was used for further comparison between two groups. Results The model group had significantly higher activities of ALT and AST than the normal group (both P < 0.000 1), and compared with the model group, the Fuzheng Huayu prescription group had a significantly lower degree of increase in ALT activity (P < 0.001). HE staining showed that the Fuzheng Huayu prescription group had a significantly lower degree of hepatocyte degeneration and necrosis compared with the model group. Compared with the normal group, the total lymphocytes, CD45, CD4^-CD8⁺ T and CD8 + CD28^-T in the model group increased significantly, while the proportion of the above lymphocytes in the Fuzheng Huayu formula group decreased significantly compared with the model group (P < 0.001). CD8⁺ T lymphocytes isolated from the liver of mice in each group were co-cultured with JS 1 for 48 hours, and compared with the control group (JS 1 cultured alone) and the normal group, the model group had a significant increase in the mRNA expression of α-SMA (both P < 0.01) and significantly higher mRNA expression of Col.I than the control group and the normal group (normal mouse liver CD8⁺ T lymphocytes co-cultured with JS 1) (both P < 0.001). The Fuzheng Huayu prescription group had significantly lower mRNA expression levels of α-SMA and Col.I than the model group (both P < 0.01). Conclusion Fuzheng Huayu prescription can indirectly inhibit activated HSCs by altering the functional phenotype of CD8⁺ T lymphocytes in mouse liver.
- Liver Cirrhosis,
- Fuzheng Huayu Recipe,
- Hepatic Stellate Cells,
- CD8-Positive T-Lymphocytes

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References

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Value of Fuzheng Huayu prescription in preventing liver fibrosis by altering the phenotypic function of CD8⁺ T lymphocytes in the liver of mice with acute liver injury

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Value of Fuzheng Huayu prescription in preventing liver fibrosis by altering the phenotypic function of CD8+ T lymphocytes in the liver of mice with acute liver injury

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Value of Fuzheng Huayu prescription in preventing liver fibrosis by altering the phenotypic function of CD8⁺ T lymphocytes in the liver of mice with acute liver injury