Cocoa procyanidins protect PC12 cells from hydrogen-peroxide-induced apoptosis by inhibiting activation of p38 MAPK and JNK

Oxidative stress induced by reactive oxygen species has been strongly associated with the pathogenesis of neurodegenerative disorders, including Alzheimer's disease. In this study, we investigated the possible protective effects of a cocoa procyanidin fraction (CPF) and procyanidin B2 (epicatechin-(4 beta-8)-epicatechin) - a major polyphenol in cocoa - against apoptosis of PC12 rat pheochromocytoma (PC12) cells induced by hydrogen peroxide (H2O2)- CPF (1 and 5 mu g/ml) and procyanidin B2 (1 and 5 mu M) reduced PC12 cell death caused by H2O2, as determined by MTT and trypan blue exclusion assays. CPF and procyanidin B2 attenuated the H2O2-induced fragmentation of nucleus and DNA in PC12 cells. Western blot data demonstrated that H2O2 induced cleavage of poly(ADP-ribose)polymerase (PARP), downregulated Bcl-X-L and Bcl-2 in PC12 cells. Pretreatment with CPF or procyanidin B2 before H2O2 treatment diminished PARP cleavage and increased Bcl-X-L and Bcl-2 expression compared with those only treated with H2O2. Activation of caspase-3 by H2O2 was inhibited by pretreatment with CPF or procyanidin B2. Furthermore, H2O2-induced rapid and significant phosphorylation of c-Jun N-terminal protein kinase (INK) and p38 mitogen-activated protein kinase (MAPK), and both of these effects were attenuated by CPF or procyanidin B2 treatment. These results suggest that the protective effects of CPF and procyanidin B2 against H2O2-induced apoptosis involve inhibiting the downregulation of Bcl-X-L and Bcl-2 expression through blocking the activation of JNK and p38 MAPK. (C) 2007 Elsevier B.V. All rights reserved.