In vitro evaluation of the protective effect of crocin on human erythrocytes

The interactions and the protective effect of the carotenoid crocin (CRO) on human erythrocytes (RBC) and molecular models of its membrane were investigated. The latter consisted of bilayers of dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylethanolamine (DMPE), representative of phospholipid classes located in the outer and inner monolayers of the RBC membrane, respectively. X-ray diffraction, differential scanning calorimetry (DSC) and electronic paramagnetic resonance spectroscopy (EPR) showed that CRO produced structural perturbations in DMPC bilayers and in isolated unsealed human erythrocyte membranes. On the other hand, scanning electron microscopy (SEM) showed that CRO induced shape changes in the RBC from their normal discoid form to echinocytes. This result indicates that the CRO molecules were mainly localized in the outer monolayer of the RBC membrane. The assessment of the protective capacity of CRO was revealed by the carotenoid inhibition of the morphological alterations caused by hypochlorous acid (HOCl) to RBC.