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Synergistic anti-inflammatory effect of gut microbiota and lithocholic acid on liver fibrosis

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Abstract

Background

Bile acids can regulate liver disease progression by affecting the functions of gut microbiota and immune cells. As the most potent natural agonist of G-protein coupled bile acid receptor 5 (TGR5) (expressed in macrophages, HSCs, and monocytes), lithocholic acid (LCA) has multiple functions, such as inhibiting inflammation and regulating metabolism. Therefore, this study aims to investigate the effects of LCA on immune cells and HSCs in liver fibrosis.

Methods

A liver fibrosis mouse model was induced by carbon tetrachloride followed by gavage of LCA, and the effects of LCA were evaluated by serum biochemical analysis, liver histology, and western bolt. Plasma cytokine levels and the number of immune cells were determined by cytometric bead array and flow cytometry, respectively.

Results

LCA could inhibit the activation of HSCs by inducing apoptosis and reducing the activation of transforming growth factor-β (TGF-β) Smad-dependent and Smad-independent pathways. Meanwhile, LCA inhibited glycolysis and promoted oxidative phosphorylation, leading to the differentiation of macrophages to M2 type and inhibiting their differentiation to M1 type. Furthermore, LCA increased the recruitment of NK cells and reduced the activation of NKT cells. However, these effects of LCA were attenuated after antibiotics reduced the diversity and abundance of the gut microbiota.

Conclusions

Gut microbiota and LCA exerted synergistic anti-inflammatory effects on liver fibrosis. The combined intervention of gut microbiota and LCA will be a new strategy for treating liver fibrosis.

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Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

AceCS1:

Acetyl-CoA synthetase

ACSL1:

Acyl-CoA synthetase

ALT:

Alanine aminotransferase

AST:

Aspartate aminotransferase

Acly:

ATP-citrate lyase

CCl4:

Carbon tetrachloride

CYP7A1:

Cholesterol 7α-hydroxylase

EMC:

Extracellular matrix

TGR5:

G protein-coupled receptors

H&E:

Hematoxylin and eosin

HSCs:

Hepatic stellate cells

IFN:

Immune interferon

Timp-1:

Inhibitor of matrix metalloproteinases 1

IL:

Interleukin

LPS:

Lipopolysaccharide

LCA:

Lithocholic acid

MMP-2:

Matrix metalloproteinase

MoMFs:

Monocyte-derived macrophages

NK:

Natural killer

NKT:

Natural killer T

NLRP3:

NOD-, LRR- and pyrin domain-containing 3

NF-κB:

Nuclear factor κB

SD:

Standard deviations

TGF:

Transforming growth factor

TNF:

Tumor necrosis factor

VM:

Vancomycin + ampicillin

α-SMA:

α-Smooth muscle actin

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Acknowledgements

We would like to express our gratitude to all those who have helped us.

Funding

The funding sources come from the National Science and Technology Major Project of China(2017ZX10202203, 2018ZX10302206).

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Authors and Affiliations

  1. State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, National Medical Center for Infectious Diseases, The First Affiliated Hospital, Zhejiang University School of Medicine, 79 Qingchun Road, Hangzhou, 310003, Zhejiang, China

    Junwei Shao, Tiantian Ge, Gang Wang & Zhi Chen

  2. Hepatobiliary and Pancreatic Surgery (Liver Disease Centre), The Second Affiliated Hospital, Zhejiang University School of Medicine, 88 Jiefang Road, Hangzhou, 310003, Zhejiang, China

    Cuilan Tang

  3. The Department of Infectious Diseases, The Second Affiliated Hospital, Zhejiang University School of Medicine, 88 Jiefang Road, Hangzhou, 310003, Zhejiang, China

    Lantian Pang

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  1. Junwei Shao
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Contributions

ZC, JS, and CT designed the experiments and wrote the manuscript. TG, JS, GW, and LP conducted the major parts of the experiments.

Corresponding author

Correspondence to Zhi Chen.

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The authors declare no competing interests.

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The authors declare that there is no conflict of interest regarding the publication of this paper.

Contribution to the field statement

Lithocholic acid (LCA) achieves anti-inflammatory and anti-fibrotic effects in the liver by regulating the differentiation of macrophages and inhibiting the activation of hepatic stellate cells (HSCs). The impacts of LCA on liver fibrosis need the synergistic effects of the gut microbiota.

Ethics approval and consent to participate

The First Affiliated Hospital of Zhejiang University School of Medicine ethics committee approved these animal experiments in accordance with Helsinki ethics committee guidelines. Informed consent was obtained from all participants. The Ethics Committee approved the experimental protocol of the same hospital (V2.0/20190503).

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Shao, J., Ge, T., Tang, C. et al. Synergistic anti-inflammatory effect of gut microbiota and lithocholic acid on liver fibrosis. Inflamm. Res. 71, 1389–1401 (2022). https://doi.org/10.1007/s00011-022-01629-4

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  • DOI: https://doi.org/10.1007/s00011-022-01629-4

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