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Journal of Physiology and Biochemistry

, Volume 72, Issue 1, pp 9–23 | Cite as

Inhibition of IRF3 expression reduces TGF-β1-induced proliferation of hepatic stellate cells

  • Ming-ming Ni
  • Tao Xu
  • Ya-rui Wang
  • Ying-hua He
  • Qun Zhou
  • Cheng Huang
  • Xiao-ming Meng
  • Jun LiEmail author
Original Paper

Abstract

Therapeutic management of liver fibrosis remains an unresolved clinical problem. Activation of hepatic stellate cell (HSC) is a pivotal event in the progression of liver fibrosis. Recent reports have showed that inhibition of activated HSC proliferation contributes to the reversal of liver fibrosis. Interferon regulatory factor 3 (IRF3), one member of the interferon regulatory factor (IRF) family, is recently proven to be a critical modulator in cardiac fibrosis. And accumulating evidence demonstrated that IRF3 plays a crucial role in liver diseases, such as hepatic steatosis, liver inflammation, and alcoholic liver injury. However, the understanding of the function of IRF3 in liver fibrosis remains limited. Our results identified the role of IRF3 in regulating human HSC (LX-2 cell) cell proliferation and apoptosis. The present study indicated that the expression of IRF3 was significantly increased in HSCs in response to TGF-β1 stimulation. Moreover, a stable and unlimited source of human HSC, the LX-2 cell line, transfected with IRF3-siRNA significantly decreases the expression level of type I collagen (Col1a1) and α-smooth muscle actin (α-SMA) in activated LX-2 cells. On the contrary, overexpression of IRF3 gives rise to an upregulation of Col1a1 and α-SMA in LX-2 cells, and further promoted HSC proliferation. Moreover, the inhibition of IRF3 significantly suppressed TGF-β1-induced HSC proliferation and increased its apoptosis. Of note, the present study indicated IRF3 may regulate LX-2 cell proliferation by via AKT signaling pathway. In summary, these observations suggest IRF3 may function as a novel regulator to modulate TGF-β1-induced LX-2 proliferation, at least in part, via AKT signaling pathway.

Keywords

IRF3 Hepatic stellate cell AKT Liver fibrosis 

Abbreviations

α-SMA

α-smooth muscle actin

DAPI

4′,6-diamidino-2-phenylindole

DMEM

Dulbecco’s Modified Eagle’s Medium

DMSO

Dimethyl sulfoxide

ECM

Extracellular matrix

FBS

Fetal bovine serum

H&E

Hematoxylin and eosin

HSC

Hepatic stellate cell

MTT

3-(4,5-dimethylthiazol-2-yl)-2,4-diphenyl-tetrazolium bromide

IRF3

Interferon regulatory factor3

PMSF

Pyrrolidine dithiocarbamate

PVDF

Polyvinylidene fluoride film

PBS

Phosphate buffered saline

SDS-PAGE

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

RT-qPCR

Quantitative real-time PCR

TGF-β1

Transforming growth factor-β1

Notes

Acknowledgments

This project was supported by grants from the National Natural Science Foundation of China (81273526, 81473268); Anhui Province Natural Science Foundation of China (1308085MH145); Anhui Science and Technology Research Projects (1301042212); and the Specialized Research Fund for the Doctoral Program of Higher Education (20123420120001)

Compliance with ethical standards

Written informed consent was provided from all of the patients, and all aspects of this study were approved by Medicine’s Ethics Committee of Anhui Medical University in accordance with the Helsinki Declaration.

Conflicts of interest

The authors declare that they have no competing of interests.

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Copyright information

© University of Navarra 2015

Authors and Affiliations

  • Ming-ming Ni
    • 1
    • 2
  • Tao Xu
    • 1
    • 2
  • Ya-rui Wang
    • 1
    • 2
  • Ying-hua He
    • 1
    • 2
  • Qun Zhou
    • 1
    • 2
  • Cheng Huang
    • 1
    • 2
  • Xiao-ming Meng
    • 1
    • 2
  • Jun Li
    • 1
    • 2
    • 3
    Email author
  1. 1.School of Pharmacy, Anhui Key Laboratory of Bioactivity of Natural ProductsAnhui Medical UniversityHefeiChina
  2. 2.Institute for Liver Diseases of Anhui Medical UniversityAnhui Medical UniversityHefeiChina
  3. 3.School of PharmacyAnhui Medical UniversityHefeiChina

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