带3蛋白磷酸化对西藏高血红蛋白献血者悬浮红细胞贮存质量的影响  

How phosphorylation of RBCs band 3 protein affects storage quality of suspended red blood cells prepared from Tibetan high Hb population

作　　者：吴小东 廖霞 李琬晶 钟锐[2] 黄敏 冯霞 张学俊[2] 王红[2] 贺曾 刘嘉馨 WU Xiaodong;LIAO Xia;LI Wanjing;ZHONG Rui;HUANG Min;FENG Xia;ZHANG Xuejun;WANG Hong;HE Zeng;LIU Jiaxin(Hospital of Chengdu Office of Peopled Government of Tibetan Autonomous Region,Chengdu 610041,China;Institute of Blood Transfusion,Chinese Academy of Medical Sciences and Peking Union Medical College)

机构地区：[1]西藏自治区人民政府驻成都办事处医院重症医学科,四川成都610041 [2]中国医学科学院北京协和医学院输血研究所

出　　处：《中国输血杂志》2022年第2期129-133,共5页Chinese Journal of Blood Transfusion

基　　金：西藏自治区2022年中央引导地方科技发展资金项目(XZ202201YD0013C);四川省医学会青年创新课题(Q15029);四川省对口科技援助项目(2021YFQ0034);西藏自治区自然科学基金(XZ2019 ZR G-127);西藏自治区科技计划项目(XZ201703-GB-02)。

摘　　要：目的分析西藏地区高血红蛋白人群血液制备的悬浮红细胞在体外保存过程中的质量变化及其临床应用的安全性。方法以体检及血液筛查合格的无偿献血者为对象,参照"青海标准"设高原高血红蛋白浓度(Hb)献血人群组(男性Hb≥210 g/L,女性Hb≥190 g/L,简称高红组):共招募西藏自治区(海拔>3 500 m)13名献血者,均为男性;对照组(男性Hb:120~185 g/L,女性Hb:115~165 g/L):共招募成都市25名献血者,其中男性12名、女性13名。采集全血200 mL/(人)袋,离心去除血浆并加入MAP保养液制备悬浮红细胞(简称悬红),分装到4个50 mL贮存袋于4℃保存。分别于保存d1、d14、d21、d35检测血常规、游离Hb、溶血率等指标,并分离红细胞膜蛋白进行带3蛋白酪氨酸磷酸化检测。结果高红组与对照组比较,RBC(×10^(12)/L)、Hct(%)、Hb(g/L)分别为6.76±0.95 vs 4.65±0.52、63.3±6.8 vs 43.1±4.4、214.4±19.8 vs 143.2±16.9(P<0.01);悬浮红细胞在贮存d1、d14、d21、d35的红细胞变形性检测:在3、5.33、9.49、16.87、30 Pa剪切力下高红组的红细胞变形性均明显低于对照组(P<0.01);高红组与对照组在贮存d1、d14、d21、d35的溶血率(%)分别为0.0502±0.0402 vs 0.0222±0.0111、0.0554±0.043 vs 0.032 1±0.0287、0.0612±0.0259 vs 0.0343±0.0317、0.0696±0.0320 vs 0.0440±0.0333(P<0.05);带3蛋白胞浆段N端Y21的磷酸化检测:在血液采集时高红组红细胞的带3蛋白呈高度磷酸化状态。结论与对照组相比,高红组悬浮红细胞的红细胞变形性降低,在体外保存过程中溶血率更高。但高红组悬浮红细胞的储存期末溶血率仍达到国家标准要求(<0.8%),高红组悬浮红细胞溶血率升高与带3蛋白的高度磷酸化密切相关。Objective To investigate the quality changes of suspended red blood cells(SRBCs) prepared from the blood of Tibetan high Hb population, and explore the availability and safety of blood collected from Tibetan high Hb population. Methods The voluntary blood donors were grouped according to the Hb concentration at the initial screening: female blood donors from Tibet Autonomous Region(>3 500 m) with Hb≥190 g/L and male blood donors with Hb≥210 g/L were classified as plateau high hemoglobin group. A total of 13 male blood donors from Tibet Autonomous Region were recruited. And the female blood donors(n=13) with Hb(115~165) g/L and male blood donors(n=12) with Hb(120~185) g/L from Chengdu were classified as control group. Whole blood of 200 mL specification was centrifuged to remove the plasma, and MAP additive solution was added to prepare SRBCs, then SRBCs were divided into four aliquots(50 mL/bag and stored at 4℃. Parameters as blood routine, free Hb and hemolysis rate were measured aseptically at day 1, 14, 21, 35 of storage. And 10 mL SRBCs was used to extract membrane proteins for tyrosine phosphorylation detection of band 3 protein. Results The RBCs counts(×10^(12)/L), hematocrit(%) and hemoglobin(g/L) of Tibetan high Hb group and control group were 6.76±0.95 vs 4.65±0.52, 63.3±6.8 vs 43.1±4.4 and 214.4±19.8 vs 143.2±16.9(P<0.01). The erythrocyte deformability test on the day 1, 14, 21, 35 of storage showed that the deformability of SRBCs prepared from Tibetan high Hb group was significantly lower than that of the control group under shear stress of 3, 5.33, 9.49, 16.87,and 30 Pa, while the hemolysis rate of SRBCs prepared from the Tibetan high Hb group and the control group on the day 1, 14, 21, 35 were 0.050 2±0.040 2 vs 0.022 2±0.011 1, 0.055 4±0.043 vs 0.032 1±0.028 7, 0.061 2±0.025 9 vs 0.034 3±0.031 7 and 0.069 6±0.032 0 vs 0.044 0±0.033 3(P<0.05). Western blotting showed that the cytoplasmic N-terminal Y21 of band 3 protein of SRBCs prepared from Tibetan high Hb group was highly phos

关 键 词：高原红细胞增多症 带3蛋白 酪氨酸磷酸化 悬浮红细胞 血液贮存 

分 类 号：R457.12[医药卫生—治疗学] Q513.3[医药卫生—临床医学]