Genistein blocks homocysteine-induced alterations in the proteome of human endothelial cells

Abstract

Dietary isoflavones from soy are suggested to protect endothelial cells from damaging effects of endothelial stressors and thereby to prevent atherosclerosis. In search of the molecular targets of isoflavone action, we analyzed the effects of the major soy isoflavone, genistein, on changes in protein expression levels induced by the endothelial stressor homocysteine (Hcy) in EA.hy 926 endothelial cells. Proteins from cells exposed for 24 h to 25 microM Hcy alone or in combination with 2.5 microM genistein were separated by two-dimensional gel electrophoresis and those with altered spot intensities were identified by peptide mass fingerprinting. Genistein reversed Hcy-induced changes of proteins involved in metabolism, detoxification, and gene regulation; and some of those effects can be linked functionally to the antiatherosclerotic properties of the soy isoflavone. Alterations of steady-state levels of cytoskeletal proteins by genistein suggested an effect on apoptosis. As a matter of fact genistein caused inhibition of Hcy-mediated apoptotic cell death as indicated by inhibition of DNA fragmentation and chromatin condensation. In conclusion, proteome analysis allows the rapid identification of cellular target proteins of genistein action in endothelial cells exposed to the endothelial stressor Hcy and therefore enables the identification of molecular pathways of its antiatherosclerotic action.

Proteomics-based Development of Biomarkers in Cardiovascular Disease

“…Two annotated 2-DE maps (26,27) have been made available. There has been work on dermal microvascular endothelial cells (28,29) and in the human endothelial cell line EA.hy926 (30,31) generated by fusion of HUVECs with the human lung carcinoma cell line A549 (32).…”

Evidence for interactions between homocysteine and genistein: insights into stroke risk and potential treatment

“…In such a way, genistein could decrease the level of free homocysteine in plasma, the effect reported previously (Chen et al 2005, Marini et al 2010. Moreover, other effects of genistein on alleviation of homocysteine-mediated changes, like modulated levels of proteins involved in metabolism and detoxification (Fuchs et al 2005), apoptosis of endothelial cells (Fuchs et al 2006), and inflammatory injury , might also be potentially explained by direct genisteinhomocysteine interactions.…”

“…In vitro studies demonstrated that genistein can reverse homocysteine-induced changes in levels of some proteins involved in metabolism and detoxification (Fuchs et al 2005). Moreover, homocysteine-dependent apoptosis of endothelial cells was inhibited by genistein (Fuchs et al 2006), and this isoflavone could protect such cells from homocysteine-caused inflammatory injury .…”

Effects of Genistein on Oxidative Injury in Endothelial Cells

“…The induction of apoptosis was associated with increased caspase-3 activity and the generation of intracellular reactive oxygen species, both of which effects were attenuated by ␣-tocopherol (22). Genistein caused inhibition of homocysteinemediated apoptotic cell death as indicated by inhibition of DNA fragmentation and chromatin condensation (23) and exhibited the strongest activity against hydrogen peroxide-induced apoptosis of HUVE-12 cells (24). The present study showed that the ox-LDL could result in a significant increase of the apoptosis index (AI) in the vein endothelia cells of oxidative injury (pϽ0.05).…”