Research of effect of astaxanthin on suspended red blood cells without leukocyte during early storage

Abstract

Objective To study the effect of astaxanthin (ASTA) on morphological structure, oxidative stress level, induding content of reactive oxygen species (ROS) and malondialdehyde (MDA), and content of 2, 3-diphosphoglycerate (DPG) in suspended red blood cells without leukocyte during early storage. Methods From May to June 2013, 6 bags of suspended red blood cells without leukocyte which were prepared at Hebei Blood Center were collected as research objects in this study by random sampling method. Then the suspended red blood cells without leukocyte of each bag were randomly divided into four groups. The ASTA was added into preservation solution of 3 groups, and the final content of ASTA was 5, 10 and 20 μmol/L, respectively, and these 3 groups were then enrolled into 5 μmol/L ASTA group, 10 μmol/L ASTA group and 20 μmol/L ASTA group. As the last group, only equivalent volume of dimethyl sulfoxide (DMSO) was added into the reservation solution, and this group was enrolled into control group. The suspended red blood cells without leukocyte of four groups were stored at 2-6 ℃. On day 7 and 14 of storage, the morphological structure of suspended red blood cells without leukocyte was observed by scanning electron microscopy. The content of ROS in suspended red blood cells without leukocyte was detected by fluorescence microplate reader. The content of MDA in suspended red blood cells without leukocyte was detected by thiobarbituric acid method. The content of 2, 3-DPG in suspended red blood cells without leukocyte was detected by ultraviolet test method. The content changes of ROS, MDA and 2, 3-DPG in suspended red blood cells without leukocyte on day 7 and 14 of storage were analyzed by statistical methods. Results ①On day 7 and 14 of storage, the percentages of double concave disc suspended red blood cells without leukocyte in 5, 10 and 20 μmol/L ASTA group were all higher than that in control group, and the percentages of suspended red blood cells without leukocyte with spines were all lower than that in control group, and all the differences were statistically significant (F=25.170, 61.376; P=0.000). The percentage of double concave disc suspended red blood cells without leukocyte in 10 μmol/L ASTA group was higher than those in 5 and 20 μmol/L ASTA group, and the percentage of suspended red blood cells without leukocyte with spines in 10 μmol/L ASTA group was lower than those in 5 and 20 μmol/L ASTA group, and all the differences were statistically significant (P 0.05). On day 7 and 14 of storage, the contents of ROS in control group were (204.3±28.7) relative fluorescence intensity/mgHb and (245.2±15.8) relative fluorescence intensity/mgHb, respectively, and both of them were the highest among four groups. And the contents of ROS in 10 μmol/L ASTA group were (148.1±5.5) relative fluorescence intensity/mgHb and (156.0±13.8) relative fluorescence intensity/mgHb, respectively, and both of them were the lowest among four groups. ③On day 7 and 14 of storage, the contents of MDA in suspended red blood cells without leukocyte of four groups were compared, and both the differences were statistically significant (F=20.698, 21.398; P=0.000). On day 7 and 14 of storage, the contents of MDA in suspended red blood cells without leukocyte of 5, 10 and 20 μmol/L ASTA group were all significantly lower than that of control group, and the content of MDA in suspended red blood cells without leukocyte of 10 μmol/L ASTA group was also significantly lower than those of 5 and 20 μmol/L ASTA group, and the content of MDA in suspended red blood cells without leukocyte of 5 μmol/L ASTA group was also significantly lower than that of 20 μmol/L ASTA group, and all the differences were statistically significant (P 0.05). On day 7 and 14 of storage, the contents of 2, 3-DPG in control group were (517.1±106.2) μg/L and (417.6±62.9) μg/L, respectively, and both of them were the lowest among four groups. And the contents of 2, 3-DPG in 10 μmol/L ASTA group were (789.1±48.2) μg/L and (677.5±90.8) μg/L, respectively, and both of them were the highest among four groups. Conclusions During early storage, ASTA could delay the change speed of morphological structure and slow down the decreasing progress of 2, 3-DPG content in suspended red blood cells without leukocyte by decreasing the oxidative stress level. But whether 10 μmol/L is the best treatment content of ASTA in suspended red blood cells without leukocyte reservation solution, it still needs further study to confirm. Key words: Erythrocytes; Astaxanthin; Reactive oxygen species; Oxidative stress; 2, 3-Diphosphoglycerate