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The Protective Effects of Helix B Surface Peptide on Experimental Acute Liver Injury Induced by Carbon Tetrachloride

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Abstract

Background

To investigate the protective effects of helix B surface peptide (HBSP) on acute liver injury induced by carbon tetrachloride (CCl4).

Methods

HBSP (8 nmol/kg) was intraperitoneally injected into C57 BL/6 mice 2 h after CCl4 administration. Serum and liver tissue samples were collected 24 h after injury. Liver function and histological injuries were evaluated. Inflammatory cell infiltration and cytokines were examined and hepatocytes apoptosis was measured. The human liver cell line LO2 and murine primary hepatocytes were stimulated by CCl4 with and without HBSP treatment and glutathione peroxidase activity, cell survival, and apoptosis were evaluated. In addition, we examined the PI3K/Akt/mTORC1 pathway to elucidate the mechanism underlying HBSP-mediated protection in acute liver injury.

Results

HBSP significantly decreased serum alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, and pro-inflammatory cytokines in liver tissues after CCl4 injection compared with those in the control group. Immunohistochemical staining indicated that the number of CD3-, CD8-, and CD68-positive cells and the expression of cleaved caspase-3 were significantly decreased by HBSP treatment. Additionally, HBSP reduced apoptosis in vivo. In an in vitro study, the glutathione peroxidase activity and survival rate increased, while the total apoptotic rate was reduced in the HBSP-treated group compared with that in the control group after CCl4 treatment. HBSP activated the PI3K/Akt/mTORC1 pathway, which was confirmed by the PI3K inhibitor LY294002 both in vivo and in vitro. Furthermore, HBSP increased the survival of mice with acute liver injury, and this effect was abolished by LY294002.

Conclusions

HBSP is a potential therapeutic agent against acute liver injury induced by CCl4.

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Abbreviations

CCl4 :

Carbon tetrachloride

TNF-α:

Tumor necrosis factor-α

IL-6:

Interleukin-6

EPO:

Erythropoietin

EPOR:

EPO receptor

βcR:

β-Common receptor

HBSP:

Helix B surface peptide

ALT:

Alanine aminotransferase

AST:

Aspartate aminotransferase

LDH:

Lactate dehydrogenase

MDA:

Malondialdehyde

MTT:

Methyl thiazolyl tetrazolium

GSH:

Glutathione

GSH-Px:

Glutathione peroxidase

qRT-PCR:

Quantitative real-time polymerase chain reaction

DMEM:

Dulbecco’s modified Eagles medium

SDS:

Sodium dodecyl sulfate

SD:

Standard deviation

SOD:

Superoxide dismutase

ANOVA:

Analysis of variance

IFN-γ:

Interferon-γ

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Acknowledgments

The study was supported by the National Natural Science Foundation of China (Nos. 81301820, 81472673, 81400752, 81401877, 81500457, 81672720), and the National Clinical Key Special Subject of China.

Author information

Author notes

  1. Shengdi Wu, Cheng Yang and Nuo Xu have contributed equally to this article.

Authors and Affiliations

  1. Department of Gastroenterology and Hepatology, Zhongshan Hospital, Fudan University, Shanghai, 200032, China

    Shengdi Wu, Yun Liu, Jiyao Wang & Xizhong Shen

  2. Department of Internal Medicine, Zhongshan Hospital, Fudan University, Shanghai, 200032, China

    Xizhong Shen

  3. Shanghai Institute of Liver Diseases, Shanghai, 200032, China

    Shengdi Wu, Jiyao Wang & Xizhong Shen

  4. Department of Urology, Zhongshan Hospital, Fudan University, Shanghai, 200032, China

    Cheng Yang

  5. Shanghai Key Laboratory of Organ Transplantation, Shanghai, 200032, China

    Cheng Yang & Lingyan Wang

  6. Department of Plastic Surgery, Zhongshan Hospital, Fudan University, Shanghai, 200032, China

    Cheng Yang

  7. Department of Respiration, Zhongshan Hospital, Fudan University, Shanghai, 200032, China

    Nuo Xu

  8. Biomedical Research Center, Zhongshan Hospital, Fudan University, Shanghai, 200032, China

    Lingyan Wang

  9. Key Laboratory of Medical Molecule Virology, Ministry of Education and Health, Shanghai, 200032, China

    Xizhong Shen

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  1. Shengdi Wu
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Contributions

SDW, CY and NX designed the research, conducted the experiments, collected and analyzed the data, and wrote the manuscript. LYW and YL contributed to the data acquisition. JYW and XZS revised the manuscript. All authors reviewed the manuscript.

Corresponding author

Correspondence to Xizhong Shen.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethics approval and consent to participate

All animal experiments were performed in accordance with the guidelines of the Care and Use of Laboratory Animals of the Laboratory Animal Ethical Commission of Fudan University with good surgical practices.

Availability of data and materials

The data and all outputs of the current study are available for testing by reviewers and scientists who wish to use them with kind full permission.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 11 kb)

10620_2017_4553_MOESM2_ESM.jpg

Supplemental Figure 1. Cytotoxicity of CCl4 at different concentrations in LO2 cells in vitro. (A) Serum ALT, AST, and MTT were measured in the groups treated with different concentrations of CCl4 (40, 60 and 80%). (B-C) Apoptosis was detected by flow cytometry analysis in the groups treated with different concentrations of CCl4. Data are represented as the mean ± SD (n = 5). (JPEG 2641 kb)

10620_2017_4553_MOESM3_ESM.jpg

Supplemental Figure  2. The expression of EPOR/βcR and Cyp2E1 in the liver. The expression of EPOR and EPOR/βcR were detected in the normal murine liver tissue (A) and murine primary hepatocyte (B). The expression of Cyp2E1 in the liver was investigated in different treatment groups (C). (JPEG 2193 kb)

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Wu, S., Yang, C., Xu, N. et al. The Protective Effects of Helix B Surface Peptide on Experimental Acute Liver Injury Induced by Carbon Tetrachloride. Dig Dis Sci 62, 1537–1549 (2017). https://doi.org/10.1007/s10620-017-4553-7

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  • DOI: https://doi.org/10.1007/s10620-017-4553-7

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