Antioxidative and protective properties of extracts from leaves of the artichoke (Cynara scolymus L.) against hydroperoxide-induced oxidative stress in cultured rat hepatocytes

Abstract

Primary rat hepatocyte cultures exposed to tert-butylhydroperoxide (t-BHP) or cumene hydroperoxide were used to assess the antioxidative and protective potential of water-soluble extracts of artichoke leaves. Both hydroperoxides stimulated the production of malondialdehyde (MDA), particularly when the cells were pretreated with diethylmaleate (DEM) in order to diminish the level of cellular glutathione (GSH). Addition of artichoke extracts did not affect basal MDA production, but prevented the hydroperoxide-induced increase of MDA formation in a concentration-dependent manner when presented simultaneously or prior to the peroxides. The effective concentrations (down to 0.001 mg/ml) were well below the cytotoxic levels of the extracts which started above 1 mg/ml. The protective potential assessed by the LDH leakage assay and the MTT assay closely paralleled the reduction in MDA production and largely prevented hepatocyte necrosis induced by the hydroperoxides. The artichoke extracts did not affect the cellular level of glutathione (GSH), but diminished the loss of total GSH and the cellular leakage of GSSG resulting from exposure to t-BHP. Chlorogenic acid and cynarin accounted for only part of the antioxidative principle of the extracts which was resistant against tryptic digestion, boiling, acidification, and other treatments, but was slightly sensitive to alkalinization. These results demonstrate that artichoke extracts have a marked antioxidative and protective potential. Primary hepatocyte cultures seem suitable for identifying the constituents responsible for these effects and for elucidating their possible mode of action.