FTIR, 1H NMR and EPR spectroscopy studies on the interaction of flavone apigenin with dipalmitoylphosphatidylcholine liposomes

Apigenin (5,7,4′-trihydroxyflavone) is a cancer chemopreventive agent and a member of the family of plant flavonoids. Apigenin interaction with liposomes formed with dipalmitoylphosphatidylcholine (DPPC) was investigated by means of FTIR spectroscopy, 1H NMR and EPR techniques. Fluorescent microscopy and electron microscopy were applied to study the apigenin effects on colon myofibroblasts and human skin fibroblasts. The strong rigidifying effect of apigenin with respect to polar head groups was concluded on the basis of the action of the flavone on partition coefficient of Tempo spin label between the water and lipid phases. The ordering effect was also found in hydrophobic region at the depth monitored by 5-SASL and 16-SASL spin labels. The inclusion of apigenin to the membrane restricted the motional freedom of polar head groups lowering penetration of Pr3 + ions to the membranes. The 1H NMR technique supported also the restriction of motional freedom of the membrane in the hydrophobic region, especially in the zone of CH2 groups of alkyl chains. FTIR analysis showed that apigenin incorporates into DPPC liposomes via hydrogen bonding between its own hydroxyl groups and lipid polar head groups in the COPOC segment. It is also very likely that hydroxyl groups of apigenin link with polar groups of DPPC by water bridges. Electron and fluorescence microscopic observations revealed changes in the internal membrane organization of the examined cells. In conclusion, the changes of the structural and dynamic properties of membranes can be crucial for processes involving tumor suppression signal transduction pathways and cell cycle regulation.

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► Apigenin incorporates into DPPC liposomes and changes their dynamic properties.
► Apigenin preferentially locates in the upper part of the membrane containing chain/glycerol/headgroup.
► Hydrogen bonds with lipid polar heads are formed by hydroxyl and keto groups of apigenin.
► COPOC segment of lipid polar head groups creates hydrogen bonds with apigenin.
► Normal biomembranes are sensitive to apigenin activity.