Ethyl caffeoate: Liquid chromatography–tandem mass spectrometric analysis in Verdicchio wine and effects on hepatic stellate cells and intracellular peroxidation

Analysis of Verdicchio (a white wine produced in Italy) with high performance liquid chromatography–tandem mass spectrometry (HPLC–ESI-MS–MS) showed a concentration of ethyl caffeoate (CfE), a natural phenolic substance, of 67.3 μmol L−1; CfE was then purified from wine with preparative reversed phase HPLC and tested on rat hepatic stellate cell cultures (HSC). HSC proliferation induced by iron(III) nitrilotriacetate (FeNTA), a pro-oxidant agent (absorbance = 0.68 ± 0.03 versus 0.32 ± 0.02 in controls), was significantly inhibited by CfE starting at a concentration of 10 μmol L−1 (absorbance = 0.45 ± 0.01), with a maximal effect at 20 μmol L−1 (absorbance = 0.32 ± 0.02). The formation of intracellular hydroperoxides induced by FeNTA was significantly reduced by CfE starting at a concentration of 10 μmol L−1 and completely prevented by CfE at a concentration of 20 μmol L−1. The increase in α1(I) procollagen mRNA synthesis induced by FeNTA was inhibited by CfE starting at a concentration of 10 μmol L−1 and almost completely at a concentration of 20 μmol L−1. Thus, Verdicchio wine is a remarkable source (about seven-fold the bioactive concentration) of CfE, an interesting drug to be tested as an antifibrotic agent.