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Protective Effect of Ganshuang Granules (肝爽颗粒) on Liver Cirrhosis by Suppressing Regulatory T Cells in Mouse Model

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Abstract

Objective

To investigate the potential antifibrotic mechanisms of Chinese medicine Ganshuang Granules (肝爽颗粒, GSG) and to provide clinical therapeutic evidence of its effects.

Methods

A cirrhotic mouse model was established by intraperitoneally injecting a mixture of CCl4 (40%) and oil (60%) at 0.2 mL per 100 g of body weight twice a week for 12 weeks. After 12-week modeling, GSG was intragastric administrated to the mice for 2 weeks, and the mice were divided into low-, medium- and high-dose groups at doses of 1, 2 and 4 g/(kg·day), respectively. Liver morphology changes were observed using Masson's trichrome staining and B-ultrasound. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and hyaluronic acid (HA) in serum were detected using an automatic biochemistry analyzer. The expressions of desmin, smooth muscle actin (SMA) and Foxp3 in liver were detected by immunoflfluorescence. The regulatory T cell (Treg) frequency was determined through flflow cytometry analysis. Collagen-I, SMA, IL-6, tumor necrosis factor α (TNF-α), interleukin (IL)-1β and transforming growth factor β1 (TGF-β1) expression levels were measured using quantitative polymerase chain reaction (qPCR).

Results

Masson's staining result showed fewer pseudolobule structures and fibrous connective tissue in the GSG-treatment groups than in the spontaneous recovery group. Ultrasonography showed that GSG treatment reduced the number of punctate hyperechoic lesions in mice cirrhotic livers. The serum ALT, AST, HA levels were significantly ameliorated by GSG treatment (ALT: F=8.104, P=0.000; AST: F=7.078, P=0.002; and HA: F=7.621, P=0.001). The expression levels of collagen-I and SMA in the cirrhotic livers were also attenuated by GSG treatment (collagen-I: F=3.938, P=0.011; SMA: F=4.115, P=0.009). Tregs, which were elevated in the fibrotic livers, were suppressed by GSG treatment (F=8.268, P=0.001). The expressions of IL-6, TNF-α and IL-1β increased, and TGF-β levels decreased in the cirrhotic livers after GSG treatment (IL-6: F=5.457, P=0.004; TNF-α: F=6.023, P=0.002; IL-1β: F=6.658, P=0.001; and TGF-β1: F=11.239, P=0.000).

Conclusions

GSG promoted the resolution/regression of cirrhosis and restored liver functions in part by suppressing Treg cell differentiation, which may be mediated by hepatic stellate cells.

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Authors and Affiliations

  1. Department of Liver Diseases Immunology, Beijing Youan Hospital, Capital Medical University, Beijing, 100069, China

    Yan-min Liu, Hong-bo Shi, Hong-lin Shi, Feng Ren & De-xi Chen

  2. Beijing Institute of Hepatology, Beijing Youan Hospital, Capital Medical University, Beijing, 100069, China

    Hong-bo Shi, Hong-lin Shi, Feng Ren & De-xi Chen

  3. Department of Toxic Hepatic Disease, Beijing Youan Hospital, Capital Medical University, Beijing, 100069, China

    Yi-rong Liu

  4. Artifical Liver Center, Beijing Youan Hospital, Capital Medical University, Beijing, 100069, China

    Yu Chen & Zhong-ping Duan

  5. Clinical Laboratory Center, Beijing Youan Hospital, Capital Medical University, Beijing, 100069, China

    Jin-li Lou

Authors
  1. Yan-min Liu
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  2. Hong-bo Shi
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  3. Yi-rong Liu
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  4. Hong-lin Shi
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  5. Feng Ren
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  6. Yu Chen
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  7. De-xi Chen
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  8. Jin-li Lou
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  9. Zhong-ping Duan
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Corresponding author

Correspondence to Zhong-ping Duan.

Additional information

Supported by the China National Key Project of Twelfth Five-Year Plan (Nos. 2012ZX10002004-006, 2012ZX10004904-003-001, and 2013ZX10002002-006-001), National Natural Science Foundation of China (No. 81300349), Beijing Natural Science Foundation (No. 7144216), Beijing Nova Program (No. Z131107000413016), and Project of Science and Technology Activities Preferred Overseas Personnel of Beijing (No. 2014)

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Liu, Ym., Shi, Hb., Liu, Yr. et al. Protective Effect of Ganshuang Granules (肝爽颗粒) on Liver Cirrhosis by Suppressing Regulatory T Cells in Mouse Model. Chin. J. Integr. Med. 25, 51–58 (2019). https://doi.org/10.1007/s11655-015-2430-9

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  • DOI: https://doi.org/10.1007/s11655-015-2430-9

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