Flow cytometric analysis of the oxidative status of normal and thalassemic red blood cells.
BACKGROUND
The oxidative status of cells has been shown to modulate various cell functions and be involved in physiological and pathological conditions, including hereditary chronic anemias, such as thalassemia. It is maintained by the balance between oxidants, such as reactive oxygen species (ROS), and antioxidants, such as reduced glutathione (GSH).


METHODS
We studied peripheral RBC derived from normal and thalassemic donors. Flow cytometric methods were used to measure (1) generation of ROS; (2) the content of reduced GSH; and (3) peroxidation of membrane lipids as an indication of membrane damage.


RESULTS
ROS and lipid peroxidation were found to be higher, and GSH lower, in thalassemic RBC compared with normal RBC, both at baseline as well as following oxidative stress, such as exposure to hydrogen peroxide. To simulate a state of iron overload, normal RBC were exposed to extracellular ferric ammonium citrate or hemin, or their Hb was denatured by phenylhydrazine. All these treatments increased ROS and lipid peroxidation and decreased GSH. These effects were reversed by N-acetyl cysteine, a known ROS scavenger.


CONCLUSIONS
Flow cytometry can be useful for measuring oxidative stress and its effects on RBC in various diseases and for studying various chemical agents as antioxidants.
Amer
Johnny
J
Department of Hematology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel.
Goldfarb
Ada
A
Fibach
Eitan
E
D004912:Erythrocytes / Q000502:physiology*; D005434:Flow Cytometry; D005978:Glutathione / Q000097:blood*; D006801:Humans; D015227:Lipid Peroxidation; D017382:Reactive Oxygen Species / Q000097:blood*; D013789:Thalassemia / Q000097:blood* / Q000473:pathology
D017382:Reactive Oxygen Species; D005978:Glutathione