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Digestive Diseases and Sciences

, Volume 62, Issue 6, pp 1537–1549 | Cite as

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

  • Shengdi Wu
  • Cheng Yang
  • Nuo Xu
  • Lingyan Wang
  • Yun Liu
  • Jiyao Wang
  • Xizhong ShenEmail author
Original Article

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.

Keywords

Helix B surface peptide Acute liver injury Inflammation Apoptosis 

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

Notes

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’s contribution

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.

Compliance with ethical standards

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.

Supplementary material

10620_2017_4553_MOESM1_ESM.docx (11 kb)
Supplementary material 1 (DOCX 11 kb)
10620_2017_4553_MOESM2_ESM.jpg (2.6 mb)
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 (2.1 mb)
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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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Shengdi Wu
    • 1
    • 3
  • Cheng Yang
    • 4
    • 5
    • 6
  • Nuo Xu
    • 7
  • Lingyan Wang
    • 5
    • 8
  • Yun Liu
    • 1
  • Jiyao Wang
    • 1
    • 3
  • Xizhong Shen
    • 1
    • 2
    • 3
    • 9
    Email author
  1. 1.Department of Gastroenterology and Hepatology, Zhongshan HospitalFudan UniversityShanghaiChina
  2. 2.Department of Internal Medicine, Zhongshan HospitalFudan UniversityShanghaiChina
  3. 3.Shanghai Institute of Liver DiseasesShanghaiChina
  4. 4.Department of Urology, Zhongshan HospitalFudan UniversityShanghaiChina
  5. 5.Shanghai Key Laboratory of Organ TransplantationShanghaiChina
  6. 6.Department of Plastic Surgery, Zhongshan HospitalFudan UniversityShanghaiChina
  7. 7.Department of Respiration, Zhongshan HospitalFudan UniversityShanghaiChina
  8. 8.Biomedical Research Center, Zhongshan HospitalFudan UniversityShanghaiChina
  9. 9.Key Laboratory of Medical Molecule VirologyMinistry of Education and HealthShanghaiChina

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