Summary
Tumor necrosis factor-alpha (TNF-α) plays a central role in cellular necrosis, apoptosis, organ failure, tissue damage, inflammation and fibrosis. These processes, occurring in liver injury, may lead to cirrhosis. Thalidomide, α-N-phthalidoglutarimide, (C13H10N2)4, has been shown to have immunomodulatory and anti-inflammatory properties, possibly mediated through its anti-TNF-α effect. In this study, we investigated the in vitro and in vivo effects of thalidomide on hepatic fibrosis. A cell line of rat hepatic stellate cells (HSC-T6) was stimulated with transforming growth factor-β1 (TGF-β1) or TNF-α. The inhibitory effects of thalidomide on the NFκB signaling cascade and fibrosis markers including α-smooth muscle actin (α-SMA) and collagen, were assessed. An in vivo therapeutic study was conducted in dimethylnitrosamine (DMN)-treated rats, which were randomly assigned to 1 of 4 groups: vehicle (0.7% carboxyl methyl cellulose, CMC), thalidomide (40 mg/kg), thalidomide (200 mg/kg), or silymarin (50 mg/kg), each given by gavage twice daily for 3 weeks starting after 1 week of DMN administration. Thalidomide (100–800 nM) concentration-dependently inhibited NFκB transcriptional activity induced by TNF-α, including IKKα expression and IκBα phosphorylation in HSC-T6 cells. In addition, thalidomide also suppressed TGF-β1-induced α-SMA expression and collagen deposition in HSC-T6 cells. Fibrosis scores of livers from DMN-treated rats receiving high dose of thalidomide (0.89±0.20) were significantly reduced in comparison with those of DMN-treated rats receiving vehicle (1.56±0.18). Hepatic collagen contents of DMN rats were also significantly reduced by either thalidomide or silymarin treatment. Immunohistochemical double staining results showed that α-SMA- and NFκB-positive cells were decreased in the livers from DMN rats receiving either thalidomide or silymarin treatment. In addition, real-time PCR analysis indicated that hepatic mRNA expressions of TGF-β1, α-SMA, collagen 1α2, TNF-α and iNOS genes were attenuated by thalidomide treatment. In conclusion, our results showed that thalidomide inhibited activation of HSC-T6 cells by TNF-α and ameliorated liver fibrosis in DMN-intoxicated rats.
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Abbreviations
- α-SMA:
-
α-smooth muscle actin
- Col 1α2:
-
collagen 1α2
- DMN:
-
dimethylnitrosamine
- G3PDH:
-
glyceraldehyde-3-phosphate dehydrogenase
- HSC:
-
hepatic stellate cell
- iNOS:
-
inducible nitric oxide synthase
- TGF-β1:
-
transforming growth factor-β1
- TNF-α:
-
tumor necrosis factor-α
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Acknowledgements
We gratefully acknowledge the kind provision of HSC-T6 cells by Dr. Scott L. Friedman, Division of Liver Diseases, The Mount Sinai School of Medicine, New York, NY, USA. We also cordially appreciate the supply of thalidomide by the TTY Biopharm Company Ltd. (Taipei, Taiwan). This work was supported in part by grants of the National Science Council, Taiwan (NSC 92-2320-077-006, NSC 93-2320-010-072 and NSC93-2320-010-001).
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Chong, LW., Hsu, YC., Chiu, YT. et al. Anti-Fibrotic Effects of Thalidomide on Hepatic Stellate Cells and Dimethylnitrosamine-Intoxicated Rats. J Biomed Sci 13, 403–418 (2006). https://doi.org/10.1007/s11373-006-9079-5
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DOI: https://doi.org/10.1007/s11373-006-9079-5