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Vitamin D and S-Farnesylthiosalicylic Acid Have a Synergistic Effect on Hepatic Stellate Cells Proliferation

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Abstract

Background

Hepatic stellate cells (HSCs) have a key role in the formation of hepatic fibrosis. The active form of vitamin D, 1,25(OH)2D3, has been found to have antiproliferative and antifibrotic effects in various tissues including liver. Farnesylthiosalicylic acid (FTS), a novel Ras antagonist, was also found to inhibit hepatic fibrosis.

Aims

The purpose of this study was to examine the antiproliferative and antifibrotic effects of the combined treatment of 1,25(OH)2D3 and FTS on primary cultured HSCs.

Methods

Primary HSCs, isolated from rat’s livers, were treated with 1,25(OH)2D3, FTS or a combination of both. Proliferation was assessed by bromodeoxyuridine. Expression of p-ERK, ERK, Ras-GTP, total-Ras, CyclinD1 and fibrotic markers was measured by western blotting analysis and real-time PCR. Cytotoxicity was assessed by lactate dehydrogenase method.

Results

The combined treatment inhibited HSCs proliferation by threefold. The effect was synergistic and non-cytotoxic. In concordance, the combined treatment suppressed CyclinD1 expression by ~2-fold, whereas 1,25(OH)2D3 or FTS alone showed a significantly lower inhibitory effect. The effect of the combined treatment on CyclinD1 expression was mediated via Ras-GTP and p-ERK signal transduction pathway. The effect on fibrotic markers showed that 1,25(OH)2D3 decreased collagen Iα1 expression by ~40 %, FTS by ~50 % and the combined treatment by ~60 %. 1,25(OH)2D3 inhibited tissue inhibitor of metalloproteinases-1 (TIMP-1) expression by 20 %. FTS alone or 1,25(OH)2D3 + FTS inhibited TIMP-1 expression by 60 %. FTS inhibited transforming growth factor-β (TGF-β) expression by 25 %, while 1,25(OH)2D3 had no effect.

Conclusion

Although the combination of 1,25(OH)2D3 and FTS did not demonstrate an additive antifibrotic effect, it showed a synergistic antiproliferative effect on primary HSCs. Therefore, the combined treatment may have a potential therapeutic value in the initiation of fibrotic process.

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Acknowledgments

This work was performed in partial fulfillment of the M.D. thesis requirements of the Sackler Faculty of Medicine, Tel Aviv University.

Conflict of interest

None.

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Authors and Affiliations

  1. Sackler Faculty of Medicine, Tel Aviv University, Tel-Aviv, Israel

    Rina Neeman & Efrat Sharvit

  2. Department of Pediatrics, Hadassah Ein-Kerem Medical Center, Kiryat Hadassah, p.o.b 12000, 91120, Jerusalem, Israel

    Shirley Abramovitch & Shimon Reif

  3. Department of Neurobiochemistry, The George S. Wise Faculty of Life Sciences, Tel-Aviv University, Tel-Aviv, Israel

    Galit Elad-Sfadia, Roni Haklai & Yoel Kloog

  4. Faculty of Medicine, Hebrew University, Jerusalem, Israel

    Shimon Reif

Authors
  1. Rina Neeman
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  2. Shirley Abramovitch
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  3. Efrat Sharvit
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  4. Galit Elad-Sfadia
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  5. Roni Haklai
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  6. Yoel Kloog
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  7. Shimon Reif
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Corresponding author

Correspondence to Shirley Abramovitch.

Additional information

Rina Neeman and Shirley Abramovitch have contributed equally to the work and should be considered “first authors.”

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Neeman, R., Abramovitch, S., Sharvit, E. et al. Vitamin D and S-Farnesylthiosalicylic Acid Have a Synergistic Effect on Hepatic Stellate Cells Proliferation. Dig Dis Sci 59, 2462–2469 (2014). https://doi.org/10.1007/s10620-014-3207-2

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  • DOI: https://doi.org/10.1007/s10620-014-3207-2

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