Biophysical characterization of genistein-membrane interaction and its correlation with biological effect on cells - the case of EYPC liposomes and human erythrocyte membranes

Bożena Pawlikowska-Pawlęga, Lucjan E. Misiak, Anna Jarosz-Wilkołazka, Barbara Zarzyka, Roman Paduch, Antoni Gawron, Wieslaw I. Gruszecki

With application of EPR and  1H NMR techniques genistein interaction  with liposomes formed with  egg yolk lecithin  and with  erythrocyte membranes was assessed. The present study addressed the problem of genistein localization and its  effects on lipid membrane  fluidity and  protein conformation. The range of microscopic techniques was employed to study genistein effects on HeLa cells and human erythrocytes. Moreover, DPPH bioassay, superoxide anion radicals test and enzymatic measurements were performed in HeLa cells subjected to genistein. The gathered results from both EPR and NMR techniques  indicated strong  ordering effect of genistein  on the motional freedom of lipids in the head group region and the adjacent hydrophobic zone in liposomal as well as in red blood cells membranes. EPR study of human ghost showed also  the changes in the erythrocyte membrane protein conformation.  The membrane effects of genistein were correlated with the changes in internal membranes arrangement of HeLa cells as it was noticed using transmission electron microscopic  and fluorescent techniques. Scanning electron  and light microscopy methods showed that   one of the aftermaths of genistein incorporation into membranes was creation of echinocytic form of the red blood cells with reduced diameter. Genistein improved redox status of HeLa  cells treated with H2O2  by lowering radicals’ level.

In conclusion, the capacity of genistein to incorporate, affect membrane organization and to change its biophysical properties is correlated with the changes inside cells.