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.
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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
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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)
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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.
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The authors declare that they have no competing of interests.
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Ni, Mm., Xu, T., Wang, Yr. et al. Inhibition of IRF3 expression reduces TGF-β1-induced proliferation of hepatic stellate cells. J Physiol Biochem 72, 9–23 (2016). https://doi.org/10.1007/s13105-015-0452-6
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DOI: https://doi.org/10.1007/s13105-015-0452-6