Adenosine A₁ receptors do not play a major role in the regulation of lipogenic gene expression in hepatocytes

Abstract

Activation of adenosine A₁ receptors was reported to promote fatty acid synthesis in AML-12 cells, by increasing the expression of SREBP-(1c) (sterol regulatory binding protein 1c) and FAS (fatty acid synthase). Since these findings have important therapeutic implications for the discovery of adenosine A₁ receptor agonists, further studies were undertaken to determine the expression and functional relevance of adenosine A₁ receptor in the liver. To that end, we used two classes of distinct adenosine A₁ receptor agonists: CPA (N⁶-cyclopentyl-adenosine), a full agonist and GS-9667 (2-{6-[((1R,2R)-2-hydroxycyclopentyl)-amino]purin-9-yl}(4S,5S,2R,3R)-5-[(2-fluorophenylthio)methyl]-oxolane-3,4-diol), a partial agonist. Treatment of AML-12 cells, HepG2 cells and primary human hepatocytes with either CPA or GS-9667 did not increase the gene expression of SREBP-(1c) or FAS. Furthermore, in AML-12 and HepG2 cells, CPA did not antagonize forskolin-stimulated cAMP production, a characteristic of adenosine A₁ receptor activation, indicating that these cells lack adenosine A₁ receptor function. Consistent with this finding, adenosine A₁ receptor gene expression was found to be very low and adenosine A₁ receptor protein levels were hardly detectable by radioligand binding assays in hepatic cell lines such as AML-12 and HepG2 as well as in both mouse and human liver tissues. Finally, acute treatment with adenosine A₁ receptor agonist GS-9667 had no significant effect on gene expression of both SREBP-(1c) and FAS in livers of Sprague Dawley rats. Taken together, our data suggest that the expression of adenosine A₁ receptor is too low to play a major role in the regulation of lipogenic gene expression in hepatocytes.