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Adenine Induces Differentiation of Rat Hepatic Stellate Cells

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Abstract

Background and Aims

Adenine is a uric acid pathway metabolite of no known function, and has recently been identified as a ligand for a rat G protein-coupled receptor. Due to the known role of other uric acid pathway metabolites in HSC biology, we tested the ability of adenine to induce HSC differentiation.

Methods

RT-PCR was performed for adenine receptor expression in T-6 and primary rat HSC. T-6 and primary rats HSC were cultured with and without adenine, and stellation examined. Next, we examined inhibition of calcium signaling using caged IP3. To test if adenine inhibits HSC chemotaxis T-6 cells and rat HSCs were cultured with or without adenine for 24 h in a transwell assay with PDGF as the chemoattractant. cDNA was prepared from T-6 and primary HSC for quantification of collagen 1 mRNA using real-time PCR.

Results

We found that mRNA for the adenine receptor is expressed in T-6 cells and primary rat HSC. Also, adenine induces HSC stellation and adenine inhibits IP3 mediated increase in cytosolic [Ca2+]i and inhibits chemotaxis in T-6 cells and primary rat HSC. Adenine was also shown to up-regulate α-SMA and collagen 1, and this effect is lost by using specific si-RNA for the adenine receptor. Finally, adenine inhibits endothelin-1-induced gel contraction.

Conclusions

The adenine receptor is present in T-6 cells and primary rats HSC. Adenine, via the adenine receptor, induces morphological change, and cytosolic calcium signaling, inhibits chemotaxis, and up-regulates collagen 1 mRNA in HSCs.

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Abbreviations

HSC:

Hepatic stellate cells

RT-PCR:

Reverse transcriptase polymerase chain reaction

TGF-β:

Transforming growth factor beta

PDGF:

Platelet derived growth factor

α-SMA:

Alpha-smooth muscle actin

MSU:

Monosodium urate

c-AMP:

Cyclic adenosine monophosphate

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Acknowledgments

This work was supported by NIH R01DK076674-01A2 and VA Merit Award (WZM).

Conflict of interest

None of the authors have a conflict of interest.

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Authors and Affiliations

  1. Section of Digestive Diseases, Yale University, 1080 LMP, PO Box. 208019, New Haven, CT, 06520-8019, USA

    Azuma Watanabe, Muhammad Adnan Sohail, Samir Gautam & Wajahat Zafar Mehal

  2. Department of Biochemistry and Immunology, Federal University of Minas Gerais, Belo Horizonte, Brazil

    Dawidson Assis Gomes

  3. West Haven VA Medical Center, West Haven, CT, USA

    Wajahat Zafar Mehal

Authors
  1. Azuma Watanabe
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  2. Muhammad Adnan Sohail
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  3. Samir Gautam
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  4. Dawidson Assis Gomes
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  5. Wajahat Zafar Mehal
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Corresponding author

Correspondence to Wajahat Zafar Mehal.

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Watanabe, A., Sohail, M.A., Gautam, S. et al. Adenine Induces Differentiation of Rat Hepatic Stellate Cells. Dig Dis Sci 57, 2371–2378 (2012). https://doi.org/10.1007/s10620-012-2183-7

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  • DOI: https://doi.org/10.1007/s10620-012-2183-7

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