NO对不同天龄红细胞保存质量影响的初步研究  

The effect of NO on the quality of RBCs in different storage condition of cell age

作　　者：于青[1] 刘嘉馨[1] 王红[1] 曹晔[1] 贺曾[1] 何语良[1] 

机构地区：[1]北京协和医学院 中国医学科学院输血研究所,四川成都610052

出　　处：《中国输血杂志》2012年第4期341-346,共6页Chinese Journal of Blood Transfusion

摘　　要：目的探讨一氧化氮(NO)对采用percoll密度梯度离心法分离的不同天龄红细胞(RBC)功能状态的影响作用。方法 1)将Percoll细胞分离液配制成5个不连续密度梯度(1.091,1.098 5,1.106,1.113 5,1.121 g/ml),将RBC按平均密度大小分层,依次吸取每层RBC分别做RBC内丙酮酸激酶(PK)含量测定并计算天龄、RBC内ATP含量检测、RBC变形性评价,以及RBC膜蛋白Western Blot。2)从同1份滤白全血中取出2等份,1份为实验组:加适宜浓度的NO溶液,另1份为对照组:加等体积红细胞保存液Ⅲ,分别于保存初期、中期、末期检测2组血液及不同天龄RBC的功能。结果 1)同1份样品经密度梯度离心分离后,RBC随着密度的增大,PK活性逐渐减小(6.192±1.25、5.165±0.84、4.538±0.76,P<0.05)、天龄逐渐增加(10.409、34.957、49.945)、ATP含量逐渐减少(4.755±0.037、3.242±0.445、2.929±0.153,P<0.01)、RBC变形性逐渐降低(200-1切变率下,0.381±0.005、0.340±0.033、0.281±0.028、P<0.05)、RBC膜带3蛋白量减小、聚簇化的带3蛋白量增多、膜上结合的IgG含量增多。2)NO组与对照组相比,全血各指标无明显差异,但NO组保存中、末期老年RBC变形性明显高于对照组(200-1切变率下,中期:0.290±0.021 vs 0.276±0.021、0.229±0.024 vs 0.211±0.027;末期:0.277±0.017 vs0.263±0.019、0.213±0.038 vs 0.193±0.039,P<0.01)。结论 RBC功能随密度的增大逐渐降低;NO可明显改善老年RBC的变形性,但对全血无明显影响。Objective To investigate the function of red blood cells （RBCs） separated by Percoll discontinuous-densi- tygradient-seperation and the effect of nitric oxide （NO）on the RBC. Methods 1 ）RBCs were separated using Percoll in saline solution corresponding to 5 different densities（ 1. 091,1. 098 5,1. 106,1.113 5,1. 121 g/ml） and removed from one layer to another; then to determinate its pyruvate kinase （PK） activity,average cell age,ATP,deformability,Western Blot of its membrane protein. 2） NO dissolved in saline solution was added to the whole blood and then to investigate its effect on the whole blood cell RBC as well as RBCs separated by according to its buoyant-density in different storage time. Results 1 ） From low to high densities, RBC PK activity （ 6. 192 ± 1.25,5.165 ± 0. 84,4. 538 ± 0. 76, P 〈 0.05 ）, ATP concentration （4.755 ±0.037,3.242 ±0.445,2.929 ±0. 153,P 〈0.01） ,the deformability（0.381±0.005,0.340 ±0.033,0.281 ± 0. 028, P 〈 0. 05 ）, the band 3 content decreased gradually, cell average age （ 10. 409,34. 957,49. 945 ）, the band 3 polymer and the binding of IgG increased. 2） There was no significant difference on the RBCs processed with NO and without NO; however,NO exerted positive effect on the RBCs with a higher buoyant-density（ mid storage period：0. 290 ± 0. 021 〉 0. 276 ± 0. 021,0. 229 ± 0. 024 〉 0. 211 ± 0. 027 ; late storage period： 0. 277 ± 0.017 〉 0. 263 ± 0. 019,0. 213 ± 0. 038 〉 0. 193 ± 0. 039, both at 200-1 shear stress, P 〈 0. 01 ）. Conclusion The function of RBC will be decreased gradually with the increase of cell buoyant-density; the addition of NO saline solution into the whole blood could potentially improve the deformability of RBC with a higher buoyant-density, but is no difference in the whole blood RBCs.

关 键 词：红细胞 密度梯度离心 红细胞保存 红细胞天龄 红细胞功能 密度 一氧化氮 变形性 

分 类 号：R331.141[医药卫生—人体生理学] R457.12[医药卫生—基础医学]