The Effects of Glutathione Monoethyl Ester on Different Biochemical, Oxidant, and Antioxidant Levels During Storage in Leukoreduced Red Blood Cells

Abstract

Background: It is essential to maintain the quality of the stored blood, because various factors affect the stored red blood cells (RBCs) over time, some red blood cell storage lesions (RCSL) develop during storage, and it could reduce the function of the RBCs. The present study aimed to evaluate the effects of glutathione monoethyl ester on different biochemical changes, oxidant, and antioxidant levels in the leukoreduced RBCs (LR-RBCs) during storage.

Materials and methods: About 10 units of LR-RBC were collected, processed and stored according to the standard operating procedures (SOPs) of the Iranian Blood Transfusion Organization. Each unit divided into 2 equal parts; LR-RBC treated with glutathione monoethyl ester and a control group. Exposure of phosphatidylserine (PS), reactive oxygen species (ROS) and microvesicle derived from the RBCs (RBC-MVs), were measured by the flow cytometry method. ELISA was used to measure the level of glutathione, and 2, 3-diphosphoglycerate (2,3-DPG). Glucose-6-phosphate dehydrogenase (G6PD) enzyme activity was measured with a chemistry autoanalyzer.

Results: The levels of glutathione reduced the initial value in the treated group (80%), and the control group (60%), respectively. Exposure of surface PS, ROS and RBC-MVs increased significantly during storage time for consecutive weeks to the amount of GSH. The levels of 2,3-DPG decreased with increasing storage time.

Conclusions: Overall, The study suggest that glutathione monoethyl ester is effective to reduce the oxidative stress and the quality of RBCs can be improved.

Keywords: Blood transfusion; glutathione; reactive oxygen species; red blood cells.

Figure 1

Identification and analysis of microvesicles = MVs,: Left side (LR-RBCs), right side (LR- RBCs + GSH). The percentage of LR-RBCs on the day 3; A) control group and B) LR-RBCs + GSH group, the percentage of RBCs-MVs on day 28; C) control group and D) LR-RBCs GSH goup, the percentage of RBCs-MVs on day 42; E) control group and F) LR-RBCs + GSH group. The population selected for 437 analysis was labeled with CD235 Phycoerythrin (PE) antibody, which was analyzed in FL2

Figure 2

The amount of PS and comparison of the effect of the glutathione monoethyl ester additive on its externalization on the surface of the RBCs during storage. In the analysis of average PS in 5 different time periods, there are significant differences with each other (P < 0.000). With comparing the effect of glutathione monoethyl ester on the amount of PS, on day 35 (P = 0.027) and 42 (P = 0.035), there was a significant difference between the two groups (P < 0.05)

Figure 3

The changes in the ROS amount level and comparison of the effect of glutathione additive on its amount during storage of the LR-RBCs. The mean percentage of the ROS in 5 different time periods has significant differences with each other (P < 0.000), compared the effect of glutathione monoethyl ester on the ROS level, except for the third day, in the rest of the study days, the difference between the two groups was significant (P < 0.05)

Figure 4

The relationship between the incidence of PS and ROS in the LR-RBCs. There is a positive correlation between the externalization of surface PS and the creation of ROS (r = 0.65 and P = 0.000)

Figure 5

Examining the relationship between the G6PD enzyme activity and the ROS. There is a negative correlation between the G6PD enzyme activity and the ROS, (r = -0.737 and P = 0.000)