Vitamin E Supplement Improves Erythrocyte Membrane
Fluidity of Thalassemia: An ESR Spin Labeling Study
Werasak Sutipornpalangkul PhD, MD*, Noppawan Phumala Morales PhD**,
Supeenun Unchern PhD**, Yupin Sanvarinda PhD**,
Udom Chantharaksri PhD**, Suthat Fucharoen MD***
Affiliation :
* Department of Orthopaedic Surgery, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand
** Department of Pharmacology, Faculty of Science, Mahidol University, Bangkok, Thailand
*** Thalassemia Research Center, Institute of Molecular Bioscience,
Mahidol University, Salaya Campus, Nakhonpathom, Thailand
Background : β-thalassemia/Hemoglobin E (β-thal/Hb E) is prevalent in Thailand. The imbalance of globin chains in red
blood cells is the primary cause of this anemic disease. The excess α-globin in β-thal/Hb E causes typical damage(s) to
membrane of erythroblasts and erythrocytes. By using three paramagnetic labeled compounds (5-, 12-, and 16-spin labeled
stearic acids, SLS), the changes of the molecular motion in the lipid bilayer of thalassemic RBCs that have structural
modification can be detected.
Objective : to investigate erythrocyte membrane fluidity and the effect of vitamin E treatment in β-thalassemia/Hemoglobin E
patients by using spin labeling techniques.
Material and Method: The erythrocyte membrane fluidity was investigated in nine splenectomized and five non-splenectomized
β-thalassemia/hemoglobin E (β-thal/Hb E) patients using EPR spin labeling techniques. To determine the effect of vitamin E
on erythrocyte membrane fluidity, only the splenectomized patients were enrolled. Patients were divided into two groups. The
first group received 350 mg vitamin E daily for a period of 1 month (n = 5) and the second group received placebo for an equal
period (n = 4). Three paramagnetic fatty acid, 5-, 12-, and 16-doxyl stearic acids, (5-, 12- and 16-DS) were used to label
phospholipids layer near both the surface (5-DS) and the deeper hydrophobic region of membrane (12-and 16-DS). Lipid
peroxidation (TBARs) was measured using a colorimetric method. Vitamin E was measured with high performance liquid
chromatography (HPLC).
Results : Significantly higher values of erythrocyte membrane fluidity were revealed with 12-, 16-DS in splenectomized
patients, as compared with non-splenectomized patients and normal subjects. In β-thal/Hb E patients, fluidity values, both
outer hyperfine splitting (2T//) and order parameter (S) of 12-DS showed inverse correlation with serum TBARs. There was
no significant difference between the fluidity values measured with 5-DS. After vitamin E supplementation, the erythrocyte
membrane fluidity was decreased in almost all patients. In contrast to the vitamin E supplementation group, increased
erythrocyte membrane fluidity was demonstrated in the placebo group. Vitamin E supplementation also had effect on other
clinical parameters such as increased plasma vitamin E, decreased serum TBARs and no change in hemoglobin.
Conclusion : The present results suggested the abnormal motion of lipid in the deeper phospholipids region of membrane. In
addition, vitamin E supplementation may have a role in the prevention of erythrocyte membrane damage of these patients.
Keywords : Thalassemia, Membrane fluidity, Vitamin E, Spin labeling
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