虾青素对悬浮红细胞储存质量的影响

The effects of astaxanthin on the storage quality of suspended red blood cells

目的观察抗氧化剂虾青素(ASTA)对悬浮红细胞储存质量的影响。方法将所采集的6名无偿献血者血液(400 m L/袋),制备成去白细胞悬浮红细胞(简称悬浮红细胞)400 m L×6袋,每袋随机均分为4组,含3个实验组:分别在去白悬浮红细胞的MAP保存液内加入ASTA使其终浓度(μmol/L)分别为5(A组)、10(B组)和20(C组);1个对照组:只加入作为ASTA溶解液的DMSO(D组)。4组悬浮红细胞于2-6℃保存至d7、d14和d35时,采用倒置荧光显微镜观察悬浮红细胞内活性氧族ROS的表达状况,以荧光酶标仪测定红细胞内ROS含量,以硫代巴比妥酸比色法测定红细胞内MDA含量,以化学比色法测定ATP含量。结果悬浮红细胞保存至d7、d14和d35时,B组ROS含量(荧光强度/mg Hb)分别为148.13±5.49、156.02±13.79、196.98±23.53,与D组相同时间点的204.26±28.69、245.17±15.81、287.52±29.02相比明显降低(P<0.05);B组MDA含量(μmol/mg Hb)分别为7.32±2.56、15.53±3.24、29.48±4.71,与D组相同时间点的23.55±5.08、33.01±4.42、50.76±7.59相比明显降低(P<0.05);B组ATP含量(μmol/g Hb)分别为7.37±1.36、5.72±0.77、4.05±0.66,与D组相同时间点的3.81±0.40、3.14±0.45、2.36±0.47相比明显升高(P<0.05)。A组和C组的ROS含量、MDA含量、ATP含量与相同时间点的D组相比,也均获得了与B组相同的结果,但其效果要逊于B组。结论 ASTA可通过抑制储存悬浮红细胞内的氧化应激水平,延缓ATP含量的降低,来改善保存质量。ASTA终浓度10μmol/L可能是保存红细胞的最适浓度。

Objective To observe the effects of antioxidant astaxanthin（ ASTA） on the storage quality of suspended red blood cells. Methods The blood from six blood donors（ 400 m L / bag） were collected to prepare suspended red blood cells in the absence of leukocytes（ 400 m L × 6）. Each bag was randomly divided into 4 groups,in which three were experimental groups. ASTA was added in the MAP preservation solution,until the final concentration of ASTA（ μmol / L） reached 5（ group A）,10（ group B） and 20（ group C）,respectively. In the control group,only the dissolved liquid DMSO of ASTA was added（ group D）. Four groups of suspended red blood cells were stored at 2- 6℃. On day 7,day 14 and day 35 of storage,the expression of reactive oxygen species（ ROS） in red blood cells was observed by inverted fluorescence microscope,The content of ROS in red blood cells was detected by Fluorescence microplate reader. The content of MDA（ malondialdehyde） was detected by thiobarbituric acid colorimetric method. The content of adenosine triphosphate（ ATP） was detected by chemical chromatometry. Results On d7,d14 and d35 of storage,the ROS content of group B（ fluorescence intensity / mg Hb） were 148. 13 ± 5. 49,156. 02 ± 13. 79,and 196. 98 ± 23. 53,respectively. Compared with the ROS content of group D（ 204. 26 ± 28. 69,245. 17 ± 15. 81,287. 52 ± 29. 02） at same points,the decrease was significant（ P〈0. 05）.On d7,d14 and d35 of storage,the MDA content of group B（ μmol / mg Hb） were 7. 32 ± 2. 56,15. 53 ± 3. 24,29. 48 ±4. 71,respectively. Compared with the MDA content of group D（ 23. 55 ± 5. 08,33. 01 ± 4. 42,50. 76 ± 7. 59） at same points,the decrease was also significant（ P〈0. 05）. On d7,d14 and d35 of storage,the ATP content of group B（ μmol /mg Hb） were 7. 37 ± 1. 36,5. 72 ± 0. 77,4. 05 ± 0. 66,respectively. Compared with the ATP content of group D（ 3. 81 ±0. 40,3. 14 ± 0. 45,2. 36 ± 0. 47） at same points,group B showed an significant increase（ P〈0. 05）. Compared with group D,the ROS,MDA and ATP content of group A and group C also achieved similar results as group B,but the effect was inferior to that of group B. Conclusion ASTA can improve the storage quality of red blood cells by inhibiting the oxidative stress level and delaying the decrease in ATP content. The final content of ASTA（ 10 mol / L） may be the most suitable content for red cell preservation.