Ethyl caffeate from Verdicchio wine: Chromatographic purification and in vivo evaluation of its antifibrotic activity

Abstract

Ethyl caffeate (CfE, caffeic acid ethyl ester) was extracted from dealcoholized Verdicchio, a white wine from Marche (Italy) with ethyl acetate and then purified with semipreparative reversed-phase high-performance liquid chromatography (RP-HPLC) using an ODS2 column (25 cm  20 mm id) at an isocratic flow of 5 mL/min (the mobile phase A was formic acid 4.5% in water and the mobile phase B was acetonitrile). The CfE extract administered intraperitoneally at 1 mmol/L in rats previously treated with 10 mg/kg dimethylnitrosamine was able to prevent the dimethylnitrosamine-induced loss in body and liver weight, as well as to reduce the degree of liver injury, as determined by alanine aminotransferase values and necroinflammatory score, after a 1-week treatment. This was associated with a reduced hepatic stellate cells activation (from 16.8 to 8.3% of smooth muscle actin positive parenchyma) and proliferation (from 11.3 to 5.5 cells/mm 2 ). The collagen synthesis was also reduced: the percentage of Sirius Red positive parenchyma decreased from 21.7 to 7.2%. The CfE levels of Verdicchio wine determined with RP-HPLC-DAD were about 14 times the active levels tested in the in vivo test. CfE can be considered as a promising natural compound for future application in chronic liver disease.

Evolution of phenolics and glutathione in Verdicchio wine obtained with maceration under reductive conditions

“…Caftaric, cis and trans-coutaric acid and fertaric are the cinnamic acids formed in the grape berries from the esterification of caffeic, coumaric and ferulic acid with tartaric acid, respectively. Verdicchio has been already reported as a good source of these congeners in previous work (Boselli et al, 2009). Since all the phenolic compounds are prone to oxidation, GSH can react with the quinonic form of the cinnamic acids through an electrophilic addition leading to the regeneration of the cinnamic form.…”

“…This compound is more lipophilic than the free form of caffeic acid (and presumably more readily absorbed by cell membranes), thus, its bioavailability is expected to be higher than non esterified phenolics such as anthocyanins or catechins. Boselli, Bendia, Di Lecce, Benedetti, and Frega (2009) have also demonstrated that CfE extracted from Verdicchio wine may exert hepatoprotective and antifibrotic effects by acting both at the cellular and molecular level in vitro and in vivo.…”

Nitrogen gas affects the quality and the phenolic profile of must obtained from vacuum-pressed white grapes

“…Caffeic acid is considered a strong antioxidant against LDL oxidation, so it contributes to the beneficial health effects of wines (Meyer, Donovan, Pearson, Waterhouse, & Frankel, 1998). Recent experimental work has evidenced that flavonols, hydroxycinnamates and their ethyl esters (Boselli, Bendia, Di Lecce, Benedetti, & Frega, 2009) are directly correlated to the antioxidant capacity of white wines (Hernanz et al, 2007;Makris et al, 2003). The knowledge accumulated so far suggests that phenols also elicit an array of biological activities, which may be associated with a depressed rate of degenerative processes (Darias-Martín et al, 2004;Fuhrman, Volkova, Suraski, & Aviram, 2001;Nardin et al, 2006).…”

Biosynthesis of ethyl caffeate via caffeoyl-CoA acyltransferase expression in Escherichia coli

“…The biological effects of L. fischeri are derived from EC. EC is known to exhibit anticancer [8], anti-inflammatory [9], and antifibrotic activities [10]. In addition, it has the potential to regulate blood pressure by inhibiting aldosterone synthase [11].…”