Abstract
Aim:
To investigate the effects of the natural product Withagulatin A on hepatic stellate cell (HSC) viability and type I procollagen production. The potential mechanism underlying the pharmacological actions was also explored.
Methods:
The effect of Withagulatin A on cell viability was evaluated in HSC and LX-2 cells using a sulforhodamine B (SRB) assay. Cell cycle distribution was analyzed using flow cytometry. Type I procollagen gene expression was determined using real-time PCR. Regulation of signaling molecules by Withagulatin A was detected using Western blotting.
Results:
Primary rat HSCs and the human hepatic stellate cell line LX-2 treated with Withagulatin A (0.625-20 μmol/L) underwent a dose-dependent decrease in cell viability, which was associated with S phase arrest and the induction of cell apoptosis. In addition, the natural product decreased phosphorylation of the Akt/mTOR/p70S6K pathway that controls cell proliferation and survival. Furthermore, Withagulatin A (1, 2 μmol/L) inhibited transforming growth factor-β (TGF-β) stimulated type I procollagen gene expression, which was attributable to the suppression of TGF-β stimulated Smad2 and Smad3 phosphorylation.
Conclusion:
Our results demonstrated that Withagulatin A potently inhibited HSC viability and type I procollagen production, thereby implying that this natural product has potential use in the development of anti-fibrogenic reagents for the treatment of hepatic fibrosis.
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Acknowledgements
We are grateful to Dr SL FRIEDMAN (Mount Sinai School of Medicine, New York) for kindly providing the human stellate cell line LX-2.
This work was supported by the National Natural Science Foundation of China (grants 20721003, 90713046), the State Key Program of Basic Research of China (grants 2009CB918502, 2010CB912501), the Shanghai Basic Research Project (09QA1406800), the Foundation of Chinese Academy of Sciences (KSCX1-YW-02-2) and the Key New Drug Creation and Manufacturing Program (2009ZX09301-001).
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Effect of Withagulatin A on procollagen I expression and Smad2/3 phosphorylation. (DOC 454 kb)
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Liu, Q., Chen, J., Wang, X. et al. Withagulatin A inhibits hepatic stellate cell viability and procollagen I production through Akt and Smad signaling pathways. Acta Pharmacol Sin 31, 944–952 (2010). https://doi.org/10.1038/aps.2010.72
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DOI: https://doi.org/10.1038/aps.2010.72
